LIMNOFISH-Journal of Limnology and Freshwater Fisheries Research 5(1): 1-5 (2019)
Thermal Tolerance of Turkish Crayfish (Astacus leptodactylus) Acclimated to Three Different Temperatures
Abdullatif ÖLÇÜLÜ 1* , Metin KUMLU 2 , H. Asuman YILMAZ 2 , O. Tufan EROLDOĞAN 2
1Fisheries Faculty, Munzur University, Tunceli, Turkey 2Fisheries Faculty, Çukurova University, Adana, Turkey
ABSTRACT ARTICLE INFO
Critical thermal maxima (CTMax) and minima (CTMin) were determined for RESEARCH ARTICLE Turkish crayfish (Astacus leptodactylus) acclimated to 15, 20 and 25°C. CTMin and CTMax were 1.3, 1.1 and 2.0°C, and 37.4, 37.5 and 38.7°C, respectively. Received : 11.05.2018 Thermal tolerance tests showed that acclimation temperatures (15, 20 and 25C) Revised : 05.10.2018 had significant effects on CTMin values of A. leptodactylus (P≤0.05). The area of Accepted : 04.11.2018 thermal tolerance assessed using the CTMin and CTMax boundaries were calculated as 364°C2. The overall ARR values were calculated as 0.07 for CTMin Published : 25.04.2019 and 0.13 for CTMax values between 15 and 25 C acclimation tempera-tures. All DOI:10.17216/LimnoFish.422903 the crayfish crumpled at 0.5°C and showed overall spasm at 32.0 – 33.0°C. Farming A. leptodactylus in the southeastern part of the Mediterranean region may be practiced in terms of temperature tolerance. * CORRESPONDING AUTHOR Keywords: A. leptodactylus, CTMin, CTMax, thermal polygon, the acclimation [email protected] response ratio Phone : +90 428 213 17 94 Üç Farklı Sıcaklığa Alıştırılan Türk Kereviti (Astacus leptodactylus)’nin Sıcaklık Toleransı Öz: 15, 20 ve 25 °C’ye alıştırılan Türk kereviti için kritik termal maksima (CTMax) ve minima (CTMin) değerleri belirlenmiştir. CTMin ve CTMax değerleri sırasıyla 1,3, 1,1 ve 2,0 ile 37,4, 37,5 ve 38,7 °C’dir. Sıcaklık tolerans testleri alıştırma sıcaklıklarının (15, 20 ve 25 °C) A. leptodactylus CTMin değerlerine önemli etkilerde bulunduklarını göstermiştir (P≤0,05). Sıcaklık tolerans alanı CTMin ve CTMax sınırları ile değerlendirilerek 364°C2 olarak hesaplanmıştır. Genel olarak 15 ile 25 C arası alıştırma sıcaklıklarında, alıştırma tepki oranı (ARR) 0,07 ile 0,13 olarak hesaplananmıştır. Kerevitlerin tamamı 0,5 °C’de kıvrılma ve geneli 32,0- 33,0°C’de kasılma göstermiştir. Akdeniz bölgesinin güneydoğu kesiminde A. leptodactylus yetiştiriciliği sıcaklık toleransı açısından uygun olabilir. Anahtar kelimeler: A. leptodactylus, CTMin, CTMax, termal poligon, alıştırma tepki oranı
How to Cite Ölçülü A, Kumlu M, Yılmaz HA, Eroldoğan OT, 2019. Thermal Tolerance of Turkish Crayfish (Astacus leptodactylus) Acclimated to Three Different Temperatures. LimnoFish. 5(1): 1-5. doi: 10.17216/LimnoFish.422903
Introduction are adaptive in determining the geographical Survival, reproduction, and growth of distribution of a species (Simčič et al. 2014). poikilothermal aquatic organisms are intimately One of the approaches to quantify lower and influenced by temperature fluctuations in their upper-temperature tolerance of aquatic animals in the environments. Ectothermic animals use a combina- laboratory is known as the critical thermal tion of behavioral and physiological mechanisms to methodology (CTM) (Cowles and Bogert, 1944). maintain their body temperatures within a narrow Thermal tolerance is quantified through the range, even under varied environmental conditions determination of the critical thermal maxima (Salas et al. 2014). The thermal preference of a (CTMax) where the temperature is increased species, therefore, corresponds closely with gradually until a critical point (lost of equilibrium) is temperatures that maximize growth and other reached (Kumlu et al. 2010a; González et al. 2010). physiological processes (Huey and Stevenson 1979; The acclimation response ratio (ARR) is an index of Diaz et al. 2011; Tepler et al. 2011). The thermal the magnitude of the thermal acclimation of an limits tolerated by an organism are set genetically and organism (Claussen 1977; Herrera et al. 1998; Díaz
2 Ölçülü et al. 2019 - LimnoFish 5(1): 1-5 et al. 2002; Pérez et al. 2003; Díaz et al. 2004; Re et was then acclimated to 15, 20 and 25°C in al. 2005). It is suggested that crustacean species that thermostatically controlled tanks at a rate of 1°C inhabit cold and temperate regions have low values increase or decrease per day. The animals were of ARR, in contrast to those from subtropical and cultured at the respective temperatures for a period of tropical regions, which present high ARR values; this 35 days prior to the onset of the experiment in round indicates that the ARR values are dependent on the fibreglass tanks (300-L) in order to stabilize their geographic gradient where the organisms are physiology and allow metabolic compensation in distributed. Attributes of thermal tolerance polygons flow through rearing systems. Each tank was fitted provide important insights into fish ecology and with 25 plastic pipes (5 cm in diameter and 10 cm distribution and have been used to identify length), which serves as hiding places for the temperature-related survival tactics (Bennett and animals. Crayfish (20-25 g) were fed twice a day Beitinger 1997). The usefulness of thermal tolerance (09.00, 17.00) by 2% of their biomass with artificial polygons lies inheritability to impart markedly more pelleted feed containing 35% protein. Water information than tolerance endpoints alone. Overall temperature was regularly checked twice a day with polygon area (reported as °C2) provides a convenient a thermometer. The food remainder, feces and moults and useful comparative index of eurythermicity were extracted daily from the reservoirs by between species (Eme and Bennett 2009). siphoning. The oxygen content and pH of rearing Turkish crayfish (A. leptodactylus) is accepted as tank was measured by an oxygen meter (OxiGuard®, a cold-water species with high economic value Birkerød, Denmark) and a WTW pH-meter (Holdich 1993; Wickins and O’C Lee 2003; (Germany). Each temperature group was then Harlioǧlu 2004). The knowledge of the critical divided into two groups to determine the CTMin and thermal minima and maxima provides a relevant CTMax in separate trials. CTMin and CTMax trials ecological index since the Turkish crayfish in nature were conducted in rectangular plastic containers (0.8 can find such spatial temperatures temporarily, as m x 0.35 m x 0.5 m) containing thermostatically acute fluctuations outside of their limit of tolerance. controlled water baths. Three containers were The CTM provides a relevant and ecological index allocated for each acclimation group and 5 crayfish when it is applied to Turkish crayfish (A. were randomly allocated to each container (n=15). leptodactylus) in its habitat. Paladino et al. (1980) During the experiment, the crayfish were not fed. mentioned that, in aquaculture, it is highly important Continuous aeration was provided for all containers to know CTMax and CTM values as they are to maintain dissolved oxygen above 5 ppm using an indicators of the thermal resistance of organisms. air blower. The photoperiod was maintained in 12 h Firkins (1993) reported thermal tolerance values for light/12 h dark. The trials were separately conducted the A. leptodactylus originating from England; at cooling or heating rates of 0.3 °C min-1. CTMin or however, basic CTM studies are necessary to CTMax was determined as the sub lethal thermal implement at different acclimation temperatures for point at which locomotory movements became A. leptodactylus originating from Turkey. Therefore, disorganized and crayfish lost the ability to escape to detect geographic influences on thermal tolerance from conditions, which ultimately lead to death. The values in crayfish, research is needed that allows for CTMin and CTMax values were calculated as the direct comparison. arithmetic mean of the collective endpoint of The aim of this study was to determine the critical individuals of a random sample of crayfish. The ARR thermal maximum and minimum, acclimation was calculated by dividing the tolerance change by response ratio (ARR) and the zone of thermal the total change in acclimation temperature according preference of Turkish crayfish acclimated to three to Claussen (1977). temperatures in order to evaluate the possibility of Statistical Analysis culturing the species in the south eastern region of the Following determination of the normality and Mediterranean of Turkey. homoscedasticity of the data, a one-way ANOVA was used to compare CTM tolerance of the species Materials and Methods (SPSS 20.1). Duncan’s multiple range test (DMRT) Critical Thermal Tolerance was carried out for post hoc mean comparisons. Sub-adult crayfish were caught in the Keban Dam Regression analysis was also carried out to evaluate Lake and transported in styrofoam boxes with wet the relationship between acclimation temperature and towels to the laboratory of Çukurova University, CTMin and CTMax (p≤0.05). Faculty of Fisheries (Adana – Turkey). The organisms were placed in a 1000-L tank, provided Results with a biological filter at 20°C for 1-week for Water quality parameters in the rearing tanks at adaptation to indoor culture conditions. Each group three acclimation temperatures were maintained at
Ölçülü et al. 2019 - LimnoFish 5(1): 1-5 3 levels appropriate for the species (Table 1). Thermal 3.0–5.0 °C, and all the crayfish crumpled at 0.5°C. In tolerance tests showed that acclimation temperatures the CTMax trials, the crayfish started to show overall (15, 20 and 25C) had significant effects on CTMin spasms at 32.0 – 33.0 °C secreting mucus like fluids values of A. leptodactylus (P≤0.05). At the at between 33.0 and 37.0 °C. The thermal tolerance heating/cooling rate of 0.3C min-1 the CTMin values polygon area was calculated as 364°C2 between 15 ranged between 1.1 and 2.0C, while CTMax values and 25°C acclimation temperatures used in the ranged between 37.4 and 38.7C (Table 2). During experiments (Figure 1). The overall ARR values were the CTMin trials, the crayfish, regardless of the calculated as 0.07 for CTMin and 0.13 for CTMax acclimation temperatures, started to lose their values between 15 and 25 C acclimation balance and laid down laterally at temperatures of temperatures (Table 3).
Table 1. Water quality parameters in the rearing tanks.
Water temperature (°C) Dissolved oxygen (ppm) pH NH3 (mg/L) 15.0±1.0 20.0±1.0 5.0-6.0 6.60-7.40 0.0 24.9±1.0
Table 2. CTMin and CTMax values of Astacus leptodactylus acclimated to 15, 20 and 25° C. Each value is a mean standart deviation (n = 15). Acclimation Temperature 15 °C 20 °C 25 °C CTMin 1.3±0.8ab 1.1±0.7b 2.0±0.8a CTMax 37.4±1.4a 37.5±0.9a 38.7±1.6a
Table 3. The calculated acclimation response ratios (ARR) for Astacus leptodactylus acclimated to 15, 20 and 25° C. Acclimation Temperature 15-20°C 20-25°C 15-25°C
ΔCTmin -0.2 0.9 0.7 ΔT 5 5 10 ARR -0.04 0.18 0.07
CTMin ΔCTmax 0.1 1.2 1.3 ΔT 5 5 10 ARR 0.02 0.24 0.13
CTMax
2004; Manush et al. 2004). Similar to the above studies, acclimation temperature was also found to significantly influence the CTMin values of A. leptodactylus in the present study. At 0.3°C min-1 cooling/heating rate, the CTMin ranged from 1.1 to 2.0 °C, and the CTMax ranged from 37.4 to 38.7 °C for crayfish reared in three acclimation temperatures from 15 to 30°C. The CTMax values obtained for A. leptodactylus were similar to those (ranged from 34.2 to 39.8 °C) reported by Díaz et al. (2002) for Macrobrachium acanthurus. A species living in the heterothermic Figure 1. Thermal tolerance polygon of freshwater environment must be tolerant to physiological crayfish A. leptodacylus over three acclimation temperature changes. The CTMax values in temperatures (15, 20 and 25°C) using CTM values. A. leptodactylus corresponded to total disorientation response and hampered locomotive activity resulting Discussion finally in death. Similar results were also reported by The importance of acclimation temperature on Hernández-Rodriguez et al. (1996) for CTMin and CTMax values has been well documented Macrobrachium tenellum, Kumlu et al. (2010a) and for various freshwater crustacean species (Layne et Kumlu et al. (2010b) for Litopenaeus vannamei. al. 1987; Espina et al. 1993; Firkins 1993; Díaz et al. During the CTMin trials, the crayfish, regardless of
4 Ölçülü et al. 2019 - LimnoFish 5(1): 1-5 the acclimation temperatures, started to lose their zones where the environmental temperatures balance and laid down laterally at temperatures of are near the final preferendum and do not exceed 5.0–3.0 °C. After the CTMin tests, almost 100% CTMax. of the animals recovered when transferred to pre-trial temperatures (Beitinger et al. 2000; Manush et al. Acknowledgements 2004). However, at CTMax tests, the crayfish started This study was financed by the Research Fund of to secrete mucus like fluids from their body at the University of Çukurova (Adana, Turkey) by temperatures of 33.0-37.0 °C. Project code number SUF2010D8 and was presented Thermal tolerance values varied within species in International Symposium on Fisheries and Aquatic with multiple estimates (Westhoff and Rosenberger Science 2016, Antalya, Turkey, 3-5 November 2016. 2016). 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A study of the factors involved in leptodactylus originating from England at four acclimatization to temperature and death at high acclimation temperatures (5, 10, 20 and 25 °C) as temperatures in Astacus pallipes. I. Experiments on 927°C2. Thermal tolerance traits could be attributed intact animals. J Cell Comp Physio. 62:119–132. to geographic and climatic differences among doi: 10.1002/jcp.1030620203 ancestral source populations in this species. Claussen DL. 1977. Thermal acclimation in On the other hand, Manush et al. (2004) calculated ambystomatid salamanders. Comp Biochem Physio thermal tolerance polygon for Macrobrachium Part A: Physio. 58(4):333–340. rosenbergii at three acclimation temperatures (25, 30 doi: 10.1016/0300-9629(77)90150-5 2 Claussen DL. 1980. Thermal acclimation in the crayfish, and 35 °C), and found a value of 255 °C , much lower Orconectes rusticus and O. virilis. Comp Biochem than that of Herrera et al. (1998), who calculated this Physio Part A: Physio. 66(3):377–384. 2 value as ˃800 °C for the same species. Díaz et al. doi: 10.1016/0300-9629(80)90183-8 (2002) reported the polygon area of 644°C2 for M. Cowles RB, Bogert CM. 1944. A preliminary study of the acanthurus at wider acclimation temperatures (20, thermal requirements of desert reptiles. Bulle Ame 23, 26, 29 and 32 °C). Mus Nat His. 83:261–296. Depending on the acclimation temperatures, the doi: 10.1086/394795 ARR values ranged from -0.04 to 0.24 for Díaz F, Sierra E, Denisse Re A, Rodríguez L. 2002. Turkish crayfish at the heating or cooling rate of Behavioural thermoregulation and critical thermal limits of Macrobrachium acanthurus (Wiegman). J 0.3 °C min-1. The overall ARR values were calculated Therm Bio. 27(5):423–428. as 0.13–0.24, which are similar to those reported for doi: 10.1016/S0306-4565(02)00011-6 some other crayfish species i.e. Astacus pallipes Díaz F, Re AD, Sierra E, Amador G. 2004. 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LIMNOFISH-Journal of Limnology and Freshwater Fisheries Research 5(1): 6-16 (2019)
Ontogenetic Diet Shift of Invasive Gibel Carp (Carassius gibelio, Bloch 1782) in Karamenderes River (Turkey)
Nurbanu PARTAL1* , Şükran YALÇIN ÖZDİLEK 2
1Çanakkale Onsekiz Mart University, Natural and Applied Sciences, 17100 Çanakkale-Turkey 2Çanakkale Onsekiz Mart University, Faculty of Arts and Science, Department of Biology, 17100 Çanakkale-Turkey
ABSTRACT ARTICLE INFO
The ontogenetic diet shift of invasive Carassius gibelio (Bloch, 1782) was RESEARCH ARTICLE investigated in Karamenderes River, Turkey. The fieldwork was performed during summer 2012, autumn 2012 and spring 2013. The fishes were caught by Received : 20.09.2018 electrofishing and using gill nets. Nine fork length groups were used in order to Revised : 10.12.2018 assess the ontogenetic diet shift. The gut contents were assessed by the index of Accepted : 12.12.2018 relative importance that was calculated from the frequency of occurrence, numerical abundance, and volumetric analyses. The most abundant length groups Published : 25.04.2019 of C. gibelio were 18-20 cm, 6-8 cm, and 27-29 cm length groups during summer 2012, autumn 2012 and spring 2013, respectively. The feeding intensity was the DOI:10.17216/LimnoFish.461758 lowest in the length groups of 15-17 cm during summer 2012, in 3-5 cm length group in autumn 2012 and in 24-26 cm length group in spring. Seasonal variations * CORRESPONDING AUTHOR were observed in the ontogenetic diet shift of C. gibelio. Large specimens [email protected] consumed more animal materials during summer and more algae in autumn. There Phone : +90 538 343 34 77 was not any significant niche overlap recorded between small and large specimens except summer. Any niche overlap between small and large specimens might be advantageous for the establishment success of invasive Gibel carp in Karamenderes River. Keywords: Gut contents, feeding intensity, diet overlap, IRI
Karamenderes Çayı’nda (Türkiye) İstilacı Gümüşi Havuz Balığının (Carassius gibelio, Bloch 1782) Beslenmesindeki Ontogenetik Değişim Öz: Bu çalışmada Karamenderes Çayı’nda bulunan istilacı Carassius gibelio (Bloch, 1782) türünün beslenmesindeki ontogenetik değişimin belirlenmesi amaçlanmıştır. Arazi çalışmaları Yaz 2012, Sonbahar 2012 ve İlkbahar 2013 mevsimlerinde gerçekleştirilmiştir. Balıkların yakalanmasında elektroşoker ile çeşitli ağlar kullanılmıştır. Beslenmedeki ontogenetik değişimi belirlemek için, balıklar çatal boylarına göre dokuz gruba ayrılmıştır. Balıkların sindirim kanalı içerikleri besin bulunma sıklığı, sayısal bolluk ve hacimsel oran kullanılarak nisbi önem indeksi ile hesaplanmıştır. C. gibelio bireylerinin mevsimlere göre bol olan boy grupları sırasıyla Yaz 2012 (18-20 cm), Sonbahar 2012 (6-8 cm) ve İlkbahar 2013 (27-29 cm) şeklindedir. Beslenme şiddeti Yaz 2012’de 15-17 cm, Sonbahar 2012’de 3-5 cm ve İlkbahar 2013’de 24-26 cm boy gruplarında en az olduğu belirlenmiştir. C. gibelio bireylerinin ontogenetik beslenme alışkanlığında zamansal ve mekânsal olarak farklılıklar gözlenmiştir. Büyük bireyler yaz mevsiminde daha çok hayvansal besin ve sonbahar mevsiminde alglerle beslendiği ve küçük bireylerle büyük bireylerin besinleri arasında yaz mevsimi haricinde önemli bir çakışma olmadığı belirlendi. Küçük ve büyük bireylerin besinleri arasında herhangi bir çakışmanın olmaması, gümüşi havuz balığının Karamenderes’de yerleşme başarısı için bir avantajı olabilir. Anahtar kelimeler: Sindirim kanalı içeriği, beslenme şiddeti, diyet çakışması, IRI
How to Cite Partal N, Yalçın Özdilek Ş, 2019. Ontogenetic Diet Shift of Invasive Gibel Carp (Carassius gibelio, Bloch 1782) in Karamenderes River (Turkey). LimnoFish. 5(1): 6-16. doi: 10.17216/LimnoFish.461758
Introduction 2011; Ekmekçi et al. 2013). In general, this species is Carassius gibelio (Bloch, 1782) is one of the known as a generalist, it has opportunistic major invasive species which was introduced to omnivorous feeding strategy and feeds on different Trace region first in the 1980s (Özuluğ et al. 2004; foods in different environments (Sakai et al. 2001; Ilhan et al. 2005) and spreaded over many freshwater Gaygusuz et al. 2006; Ekmekçi et al. 2013). It is systems rapidly throughout Turkey (Aydın et al. obvious that high variety in food resources of this
Partal and Yalçın Özdilek 2019 - LimnoFish 5(1): 6-16 7 invasive species will affect many other indigenous has such advantage make it successive in establish species living in the same habitat (Goodell et al. and spread. 2000) and make it more advantageous among competitors. In addition to having advantages of Materials and Methods this species in the interspecific relationship, Study area and sampling the high variety of food resources may change during Karamenderes River, about 109 km in length, ontogeny and the intraspecific resource partitioning originates from the Kaz and Ağı Mountains and may another advantage of this species directs to West and North and flows into Çanakkale in the introduced ecosystems. There are many studies strait after watering Kumkale plate in Biga Peninsula, about ecological traits (Lockwood et al. 2013; Çanakkale. There are two reservoirs along the river. Ekmekçi et al. 2013; Tarkan 2013), gut contents One is in the Bayramiç province, which is about 86.5 (Specziár et al. 1997; Rybczyk 2006; Yılmaz et al. cubic hectometer water capacity, and the other is in 2008; Rogozin et al. 2011; Partal 2014; Partal and Pınarbaşı village, which is smaller than the other. Yalçın Özdilek 2017) and feeding characteristics These reservoirs are used for irrigation purposes (Specziár et al. 1997; Rybczyk 2006; Yılmaz et al. (Figure 1). 2008; Rogozin et al. 2011; Partal 2014; Yalçın C. gibelio is first recorded in Karamenderes at the Özdilek and Jones 2014; Partal and Yalçın Özdilek lower part of the Pınarbaşı village after fish stocking 2017) of this species. C. gibelio has been first studies on Bayramiç Dams by the activities of recorded in Karamenderes river which is Directorate of National Water Affairs (Yalçın on the Northwestern part of Turkey in 2007 (Yalçın Özdilek 2008). The samplings were conducted in Özdilek 2008). There are some records three seasons, during summer 2012 (July-August on the feeding habits of C. gibelio from 2012), autumn 2012 (October-November 2012), and Karamenderes river (Yalçın Özdilek and Jones 2014; spring 2013 (May 2013). Sampling could not be Partal and Yalçın Özdilek 2017), however, there is a performed in winter because of inconvenient weather gap in the knowledge about ontogenetic diet shift of conditions for sampling. The fish sampling has this species. There is a limited study on the performed at 14 stations along the Karamenderes ontogenetic diet shift of C. gibelio and the data on River from the upper parts of the dams to the river this subject with the ontogenetic niche overlap mouth. The names of the stations from up to down and trophic position will serve to understand the are Karaköy 1, Karaköy 2, Evciler, Evciler trout establishment success of this species. farm, Çırpılar, Mollahasanlar, Bayramiç-Çan road, The morphological, physiological and behavioral Ahmetçeli, Sarmısaklı, Pınarbaşı, Kalafat, Kumkale changes during the developmental stage may result in bridge (3), Kumkale closed end (2), Kumkale open ontogenetic diet shift (Wilbur 1980; Miller and end (1) (Figure 1). Rudolf 2011; De Roos and Persson 2013; Nakazawa Different sampling device was used for fishing 2015). In addition, changes in foraging according to habitat characteristics. Fish were ability depending on the growth may cause the collected by scanning about 20 m lengths of the river ontogenetic diet shift in fish (Bergman and during 20 minutes in every station by electrofishing Greenberg 1994; Jeppesen et al. 2003; Alcaraz and (SAMUS) on the upper sites of Karamenderes. Gill García-Berthou 2007; Nakazawa 2015). Shifting in nets in different mesh size (18 mm, 22 mm, 25 mm, feeding pattern is common with a function of age and 32 mm knot to knot) and different lengths (160 and length in many animal species (Wilbur 1980; m-2.5 m, 100 m-2 m, 15 m-2 m, and 30 m-1 m) were Miller and Rudolf 2011; De Roos and Persson 2013; used for fishing (average of 24 hours) in the river Nakazawa 2015). Data on the ontogenetic diet mouth stations. In addition, fyke net composed of 8- shift is very important for evaluating the ecological 37 sets each has 12 m long and a cast net, which has role of a species (Werner and Gilliam 1984; 10 mm mesh size and its radius 140cm were used for Post 2003). Intraspecific competition reduces the fishing in some stations. All the fish samples were population growth particularly in the limited transferred to the laboratory in an icebox and after resource condition (Bolnick et al. 2011). The data on identification, they preserved in a deep freeze with the ontogenetic diet shift of invasive C. gibelio labeled. may serve to take some measures related to mitigating the adverse effects of this Laboratory procedures invasive species. We aimed to reveal the The gut contents of 215 specimens of totally 251 food diversity in different length groups and specimens could be examined. Before dissection, the the ontogenetic diet shift of C. gibelio fork length and weight were measured by the quantitatively in this study. This study standard ruler (± 0.01 mm) and a balance (± 0.1g). will serve to understand if this species After dissection, the total gut length from the
8 Partal and Yalçın Özdilek 2019 - LimnoFish 5(1): 6-16 esophagus to the anus was measured using the same IRI = (N %+V %) x F % ruler. The sex of the specimens was determined under Fi %= i prey items frequency of occurrence in the a stereomicroscope. The gut contents were evacuated gut x 100 / total number of full guts in a graduated cylinder, which contains 70 % ethanol. N %= i prey items total number x 100 / prey items The total gut volumes were measured by the i total number replacement of ethanol level. Diluted gut contents in a Sedgewick- Rafter lam Vi %= i prey items total volume x 100 / prey items were examined under a stereomicroscope x10 total volume magnitude. The number and sizes (Sun and Liu 2003) of each food category were recorded after the 퐻 = − ∑ 푝푖푙푛푝푖 description of the taxon at the possible level. The percentage of empty guts, vacuity index (VI 푝푖 = 푁푖⁄푁 %), were used to assess the feeding intensity (Hureau equation. The trophic level was calculated as 1966). The feeding intensity was assessed taking into consideration the length groups, sex, season and 11 stations. Vacuity Index (VI) was used for calculating 푇퐿푘 = 1 + (∑ 푃푗 × 푇퐿푗) the feeding intensity by using 푗=1 VI = empty gut number x 100 / total gut number in the equation (Cortés 1999). The IRI % values of equation (Hureau 1966; Costa and Cabral 1999). each food category were used to calculate the The percentage of the relative index (IRI %) was diversity and trophic position. evaluated to assess gut contents data using the Fish were grouped into nine-length class frequency of occurrence (F %), numerical (N %) and categories and the differences in IRI % value of each volumetric (V %) methods (Pinkas et al. 1971; Prince food category in each length group were tested by 1975; Hyslop 1980). nonparametric Kruskal Wallis test.
Figure 1. Sampling area (Partal and Yalçın Özdilek 2017 (adapted)). Results dam. 62 %, 27 % and 12 % of the specimens were A total of 215 specimens were all caught at the collected during summer 2012, autumn 2012 and lower stations of Bayramiç Reservoir. There were no spring 2013, respectively. The spatial and seasonal specimens encountered at the upper stations of this relative abundance of specimens indicates that
Partal and Yalçın Özdilek 2019 - LimnoFish 5(1): 6-16 9 the most abundant specimens were recorded at length group was 6-8 cm FL with the percentage of the Kumkale River mouth station in summer, 16.7 in total. The 18-20 cm, 6-8 cm, and 27-29 cm FL Ahmetçeli station in autumn and Kumkale bridge (3) groups were the most abundant length groups in station in Spring with the percentages of 60.9, 14.9 summer, autumn and spring seasons, respectively and 16.1, respectively (Figure 2). The most abundant (Figure 3).
Figure 2. The distribution and relative abundance of C. gibelio specimens according to seasons and the station.
Figure 3. The distribution and relative abundance of the length groups according to the seasons. Table 1. Shannon-Wiener Diversity Index (SWI) and Vacuity Index (VI) values according to the length groups. Length Groups (cm) Sampling 3-5 6-8 9-11 12-14 15-17 18-20 21-23 24-26 27-29 Season SWI VI SWI VI SWI VI SWI VI SWI VI SWI VI SWI VI SWI VI SWI VI Summer 2012 2.23 9.1 2.51 0 2.34 20 2.46 33.3 1.76 55.6 2.26 37.5 1.86 34.8 1.39 44 - - Autumn 2012 1.69 33.3 2.05 0 2.23 23.1 1.78 20 1.56 14.3 0.89 0 1.7 - 1.47 0 1.57 0 Spring 2013 - - 1.39 0 - - 1.41 0 2.19 16.7 1.64 0 2.04 0 1.99 25 1.63 0
10 Partal and Yalçın Özdilek 2019 - LimnoFish 5(1): 6-16
Gut Contents Oligochaeta setae in nearly every size group in every The gut contents of C. gibelio consisted of season. siliceous algae, green algae, vascular plants, pine When the gut contents were grouped as four-diet pollen, amphipods and chironomids (Table 2). Algae categories (detritus, periphyton, macrophyte, and took an important part in the gut contents by macroinvertebrate) periphyton dominate nearly all frequency and abundance. As members of length groups except 15-20 cm length groups in Bacillariophyceae, Navicula sp., Fragilaria sp. autumn according to N% values (Figure 4). and partly Cocconeis sp. taxa were dominant Particularly, macroinvertebrates were low values organisms in nearly all gut contents. Some animal both abundance and volume in > 20 cm length groups such as Oligochaeta members could not specimens in autumn. The V% values of include in the gut content analysis due to rapid macroinvertebrates were low in <15cm and >20 cm digestion. However, there were encountered specimens in spring and autumn, respectively.
100% 100% 100% 90% 90% 90% 80% 80% 80% 70% 70% 70% 60% 60% 60%
50% 50% 50%
% 40% 40%
40%
F 30% 30% 30% 20% 20% 20% 10% 10% 10% 0% 0% 0%
a b c
100% 100% 100%
80% 80% 80%
60% 60% 60%
40% 40%
% 40%
N 20% 20% 20%
0% 0% 0%
a b c
100% 100% 100%
80% 80% 80%
60% 60% 60%
%
V 40% 40% 40%
20% 20% 20%
0% 0% 0%
a b c
100% 100% 100%
80% 80% 80%
60% 60% 60%
%
IRI 40% 40% 40%
20% 20% 20%
0% 0% 0%
a b c
Fifure 4. The occurrence (F %), number (N % ), volume (V %) and Index of Relative Importance (IRI %) of prey items in the gut contents, in terms of the length groups and seasons (a: Summer 2012; b: Autumn 2012; c: Spring 2013. Blue: detritus; red: periphyton; green: macrophyte; purple: macroinvertebrate)
Partal and Yalçın Özdilek 2019 - LimnoFish 5(1): 6-16 11
Table 2. Seasonal (SUM: Summer 2012; A: Autumn 2012; SP: Spring 2013) IRI % values of length groups.
Seasonal IRI % Values of Length Groups 3-5 6-8 9-11 12-14 Prey Items SUM A SUM A SP SUM A SUM A SP Cyanobacteria Oscillatoria sp. 1.82 14.14 2.33 Amphora sp. 1.19 0.33 1.29 0.23 2.34 2.70 0.40 0.80 Cocconeis sp. 0.34 3.90 1.34 2.0 7.30 10.25 2.50 10.45 2.8 Cyclotella sp. 0.19 Cymatopleura sp. 0.17 0.74 0.08 0.10 0.45 Cymbella sp. 0.22 5.26 0.19 0.7 15.65 1.17 5.54 3.52 Diatoma sp. 0.04 0.09 0.06 0.01 Fragilaria sp. 0.01 0.91 0.07 8.64 0.08 0.07 0.16 Fragilaria sp. (chain) 13.48 0.54 17.49 1.26 16.8 2.37 15.04 18.58 0.24 54.8 Gomphonema sp. 0.35 1.95 0.12 3.96 2.18 0.13 Gyrosigma sp. 0.34 0.40 0.25 0.07 0.7 12.07 1.20 4.18 0.66 Licmophora sp. 1.44 6.70 1.10 Melosira sp. 5.82 8.79 0.30 12.49 0.93 5.95 2.44
Heterokontophyta Navicula sp. 8.98 41.69 11.54 43.19 15.4 16.94 28.45 6.71 31.54 6.1 Neidium sp. 0.06 Nitzschia sp. 2.88 1.73 16.55 0.15 Pinnularia sp. 2.34 0.004 0.76 0.05 0.20 0.16 Rhoicosphaenia sp. 2.28 0.33 0.94 0.40 0.07 0.05 0.11 Stephanodiscus sp. 0.03 0.35 0.41 0.003 1.23 0.16 Ulnaria sp. 0.95 1.10 0.07 10.7 0.02 0.26 0.66 Vaucheria sp. 0.31 Ankistrodesmus sp. 0.6 Chlorella sp. 1.53 Chlorophyta 0.03 Cladophora sp. 1.10 2.06 0.56 2.70 0.46 6.80 Closterium sp. 0.19 0.06 0.15 0.8 0.70 Conjugatophyceae
Cosmarium sp. 0.02 0.02 0.02 Microspora sp. Oedogonium sp. 0.01 0.19 0.05 3.51 0.03 0.16 0.25
Chlorophyta Pandorina sp. Pediastrum sp. 0.10 0.04 1.08 Scenedesmus sp. 0.01 0.02 0.06 Spirogyra sp. 5.78 1.002 Stigeoclonium sp. 0.39 0.73 0.06 0.99 0.33 0.61 0.38 0.7 Ulothrix sp. 0.01 Zygnema sp. 0.14 Amphipoda 23.94 7.99 5.79 1.49 18.23 19.50 9.73 Chironomidae 0.11 52.3 0.25 Copepoda 11.04 0.56 Crustacea 0.29 Insecta 3.91 2.46
Keratella sp. 0.46 Nematoda Ostracoda 1.94 0.53 6.21 2.72 8.31 Plant (seed) 0.01 Pollen 0.20 0.27 0.04 0.004 0.02 Plant 0.23 10.15 7.59 0.13 2.12 0.13 11.98 14.8 Fish egg 0.04 0.10 0.04 0.03 3.6 Animal and Plant Prey and Plant Animal Animal Detritus 15.78 19.07 0.80 15.5 Digested Detritus 0.41 0.02 0.32 Other organisms 16.58 25.71 4.41 2.39 0.7 2.44 4.37 8.97 0.13 1.2 Cystic material 0.001 0.02 Bryozoa 0.03 0.38 2.82 30.87 Cladocera 0.10
12 Partal and Yalçın Özdilek 2019 - LimnoFish 5(1): 6-16
Table 2. (Continous)
Seasonal IRI % Values of Length Groups 15-17 18-20 21-23 24-26 27-29 Prey Items SUM A SP SUM A SP SUM A SP SUM A SP A SP Anabaena sp. 0.003 Cyano- Merismopedia sp. 0.02 bacteria Oscillatoria sp. 0.001 1.28 0.002 Amphora sp. 3.94 3.65 0.02 3.29 1.82 2.35 0.07 0.01 0.17 0.1 Cocconeis sp. 1.49 10.53 0.29 7.03 3.64 3.1 6.96 5.18 2.72 2.66 5.96 0.5 0.5 Cyclotella sp. 0.01 Cymatopleura sp. 0.09 0.03 0.32 0.03 0.2 0.15 0.42 0.01 0.23 Cymbella sp. 0.34 1.22 2.02 6.38 3.09 0.27 0.37 0.23 12.8 1.48 0.3 Diatoma sp. 0.01 0.36 0.003 0.01 0.001 Epithemia sp. 0.07
Fragilaria sp. 0.09 0.33 0.14 0.001 Fragilaria sp. (chain) 0.04 1.51 28 3.11 1.84 45.5 0.76 6.38 37.5 5.02 4.32 6.4 1.22 21.7 Gomphonema sp. 0.01 0.21 0.08 0.04 0.11 0.02 Gyrosigma sp. 2.15 0.28 1.65 2.56 2.28 1.04 0.06 0.79 1.25 Melosira sp. 0.49 0.84 8.73 2.85 1.63 16.39 5.36 0.93 34.9 4.9 27.8 37.3 Meridion sp. 0.004 0.003 Heterokontophyta Navicula sp. 8.7 3.34 16.6 5.32 1.82 15.3 6.34 44.46 0.86 1.12 38.2 6.4 26.1 3.2 Nitzschia sp. 1.32 0.03 0.76 1.31 0.29 0.02 Pinnularia sp. 0.01 0.11 0.02 0.003 0.002 0.03 Rhizosolenia sp. 0.0001 Rhoicosphaenia sp. 0.72 0.72 0.10 1.82 2.03 0.39 0.87 0.06 Stephanodiscus sp. 0.13 0.01 0.60 0.03 0.01 1.6 0.2 Surirella sp. 0.001 Ulnaria sp. 5.46 5.90 2.80 3.5 1.66 0.16 0.38 1.95 2.5 2.3 Ankistrodesmus sp. 0.003 0.001 Chaetomorpha sp. 0.02 Chlorophyta 0.03 0.01 Cladophora sp. 0.01 1.04 2.22 3.35 0.54 Closterium sp. 0.003 0.03 0.01 0.02 Conjugatophyceae 0.10 0.26 9.26 0.01 7.3 0.2
Cosmarium sp. 0.01 0.09 0.003 0.01 Microspora sp. 0.13 0.13 0.08 0.07 Oedogonium sp. 0.01 0.51 0.05 15.81 0.66 0.01 3.4 1.65 Pandorina sp. 0.002
Chlorophyta Pediastrum sp. 0.02 0.005 0.15 Scenedesmus sp. 0.03 0.001 0.001 0.0004 Spirogyra sp. 0.09 0.01 3.29 1.4 Stigeoclonium sp. 0.02 0.08 0.24 0.07 0.18 0.03 0.21 Ulothrix sp. 0.35 0.003 Ulvales 0.05 Zygnema sp. 0.01 0.15 Charophyta Mougetia sp. 0.01 Amphipoda 51.62 38.70 0.38 21.45 76.1 0.9 52.42 4.04 0.46 59.97 2.003 13.9 0.22 27.5 Chironomidae 0.91 0.0003 2.26 1.34 21.3 0.25 4.87 1.8 Copepoda 0.52 0.0003 0.27 2.02 0.02 0.27 0.10 Crustacea 0.12 0.11 0.18 0.80 3.83 Gastropoda 0.001 0.01 0.08
Insecta 0.03 0.26 0.36 0.38 0.20 9.35 0.55 0.5 Nematoda 0.003 0.0001 Ostracoda 16 36.47 34.48 12.9 13.79 20.89 1.03 Pollen 0.005 0.01 0.11 0.1 0.17 0.25 1.12 Plant 3.80 0.14 10.1 0.38 1.4 0.95 13.7 0.39 42.8 0.98 Fish egg 0.45 3.65 0.28 0.7 0.08 10.9 0.47 3.96 0.11 0.04 Animal Detritus 0.01 1.67 0.002 0.001 3.7
Animal and Plant Prey and Plant Animal Digested Detritus 0.77 0.07 1.97 0.23 1.19 0.14 1.6 Other organisms 0.59 1.09 15.5 1.58 3.9 0.86 4.95 0.15 0.20 0.9 1.02 1.1 Plant (circle shaped) 0.06 0.03 0.001 0.06 Cystic material 0.23 0.01 0.02 0.12 Bryozoa 0.56 33.5
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Ontogenetic diet shifts Amphipoda members were 23.9 %, 18.2 %, and The ontogenetic diet shift indicated seasonal Copepoda members 11.0 % in autumn, variation taking into consideration IRI % Chironomidae members were high (52.3 %) values. There was a significant difference among in spring in smaller than 12 cm FL group (Table 2). IRI % of various length groups in all three seasons (X=27.003, P<0.01, df=7; X=20.603, P<0.05, Relative gut length and feeding strategy df= 8; X=14.073, P<0.05, df= 6). Heterekontophyta The mean relative gut length of C. gibelio is members were the most common food groups 3.47 ± 0.85 with the range of 1.22-5.66 (in summer) in nearly all length groups. While the small 2.60 ± 0.73 with the range of 1.03-3.84 (in autumn) specimens (3-4 cm) feed on mostly 3.52 ± 0.56 with the range of 2.12-4.56 (in spring). C. Heterokontophyta members, the larger gibelio specimens have omnivore feeding strategy specimens consumed mostly animal foods such as according to their length groups. (Table 3). When Amphipoda and Ostracoda in all the seasons. group’s specimens that were smaller than 12.0 cm Particularly, the IRI % value of Amphipoda members length collected in three seasons combined, the mean was more than 50 % in the large specimens in RGI value of this combined group was 2.51 ± 0.77 summer. Navicula sp. was the highest IRI % in the that means carnivorous dominant omnivorous 3-11 cm FL group in autumn. According to three feeding strategy. The RGI values of 12.1-17.9 cm and season data, the critical length group shifting diet larger than 18.0 cm length groups combined in three from the herbivorous to carnivore dominated feeding seasons were 3.38 ± 0.67 and 3.66 ± 0.69 strategy is 12 cm FL. However, the IRI % of respectively. Table 1. Taking into consideration RGI types of feeding in different length groups (O: Omnivorous; H: Herbivorous; C: Carnivorous; N: Number of specimens; GL: Gut length; FL: Fork length; SD: Standart Deviation). Length groups (cm) N Summer (GL/FL±SD) N Autumn (GL/FL±SD) N Spring (GL/FL±SD) 3-5 11 2.47±0.97 O-C 3 1.55±0.34 C-O - - - 6-8 10 2.69±0.43 O 25 2.47±0.81 C-O 1 3.16 O 9-11 5 3.15±0.92 O 13 2.40±0.56 C-O - - - 12-14 6 3.65±0.75 O 5 2.85±0.36 O 1 3.13 O 15-17 20 3.52±0.73 O 7 3.24±0.26 O 6 3.21±0.79 O 18-20 23 3.69±0.74 O-H 1 3.33 O 2 3.67±0.95 H-O 21-23 22 3.70±0.78 O-H 3 3.01±0.38 O 4 3.58±0.71 O 24-26 25 3.73±0.76 O-H 1 3.04 O 4 3.81±0.36 H-O 27-29 - - - 1 3.62 O 7 3.64±0.19 O
Trophic level the spring and summer seasons. The trophic level of The trophic level of all C. gibelio specimens larger than 20 cm FL specimens decreased ranged from 2.03 to 3.34. The trophic level was dramatically in autumn. The minimum and maximum low and more or less steady state in spring comparing TLs of all groups are 2.06-3.29, 2.03-3.34 and 2.07- to summer and autumn. The trophic level is 2.79 in summer, autumn, and spring, respectively increasing at larger than 12 cm FL specimens both in (Figure 5).
4.00
3.50
3.00
2.50
2.00 Summer 2012 Autumn 2012
TrophicLevel 1.50 Spring 2013 1.00
0.50
0.00 3-5 6-8 9-11 12-14 15-17 18-20 21-23 24-26 27-29 Length groups, cm
Figure 4. Trophic level of the length groups.
14 Partal and Yalçın Özdilek 2019 - LimnoFish 5(1): 6-16
Discussion intensity reported by Kırankaya (2007) and Bobori et C. gibelio were observed only the lower part of al. (2012). In general, the feeding intensity estimated the Bayramiç Dam along the River Karamenderes. based on a number of empty gut indicates seasonal The spatial distribution of C. gibelio had seasonal variation, with low in hot summer season because of variation along the River Karamenderes. While the increasing enzyme activities and digestion specimens larger than 13.6 cm FL were abundant at metabolism. The seasonal and size depend variation the river mouth station, the specimens smaller than in the feeding intensity might be related with the diet 8.8 cm FL were rich at the upper sites just below the types. For example, in summer the specimens smaller Bayramiç dam in the summer and spring. The smaller than 8 cm TL feed mostly on plant materials and specimens might escape from the Bayramiç Dam, likely in autumn the large specimens consumed plant which is regularly fished by aquaculture activities material. It is important that the digestion of plant and the most available habitat for C. gibelio might be material is hard when comparing to animal material at the river mouth conditions. and the retention duration of plant materials is longer A previous study based on a gravimetric method than that of animal materials in the gut (Nikolsky indicates that small specimens mostly feed on 1978). In addition, the smaller specimens might feed diatoms and large specimens consume animal on relatively small food and the retention duration of materials such as copepods, beetles and chironomids small animal materials as a diet would be relatively in a Brackish Water Body in Southern Siberia shorter time (Labropoulou et al. 1997) in gut content (Rogozin et al. 2011). Another study, which is based comparing to large animal materials which are on F % and N % indicates that small specimens presumably consumed by larger specimens. In other consume zooplanktonic organisms such as copepods words, the feeding intensity might be explained by and cladocerans in Gelingüllü reservoir, Turkey fish abundance. In small and large specimens was (Kırankaya 2007). In addition, the phytoplankton was low abundance in summer and autumn, respectively recorded as low frequency comparing to other food (Figure 3). This indicates that when the population of organisms in all age groups in Gelingüllü reservoir C. gibelio low density, they prefer the most available (Kırankaya 2007). There are no previous study and abundant plant materials as food. The small recording gut contents as IRI % values, however, the specimens mostly feed on siliceous algae and results obtained from abundance and frequency in probably feed on the mats of periphyton, which are Karamenderes River were apart from the results of more abundant in the shallow, pooled, high vegetated the previous records. Periphyton was important diet stony and macrophytes dominant microhabitats along as frequency and abundance, as well as IRI %, in the river. This kind of habitat might be more suitable almost all length groups. for small specimens serve them both food and shelter The amount of animal food might be enough only for escaping their predators. Yalçın Özdilek and large specimens who are more capable to collect the Jones (2014) stated that the filamentous algae were animal materials comparing to smaller ones, so the important food components for about middle size smaller ones have to change their feeding (13.5-21.1 cm in FL) C. gibelio living in characteristics into algae because of competition. Karamenderes. However, IRI % results indicate that This finding was supported by our high diet diversity 12-21 cm length group members feed on animal prey in the gut contents of smaller specimens in items in overall stations in Karamenderes river in this Karamenderes River. There is no diet diversity in C. study. This study supports that C. gibelio is an gibelio feeding patterns in the previous study, and our opportunistic feeder and the plasticity in its feeding results indicated a decrease in Shannon diversity in strategy might be seasonal and ontogenetic. food components of >12 cm C. gibelio. This C. gibelio has an omnivorous feeding strategy as indicated that gibel carp has a wide plasticity in every indicated many studies (Specziár et al. 1997; Balık et length group and this wide range on the capability of al. 2003; Rybczyk 2006; Kırankaya 2007; Yılmaz et resource use give them a high advantage for al. 2008; Yalçın Özdilek and Jones 2014; Partal and surviving in even very limited resource conditions. In Yalçın Özdilek 2017). The trophic level based on IRI the other views, the food selection of specimens % and RGI data support this finding. The ontogenetic might be related to the abundance of resource users. diet patterns taking into consideration, there was a While the abundance of large Gibel carp specimens conflict between IRI % and RGI results. Taking into were abundant in summer, the small ones were consideration RGI carnivorous dominant feeding abundant in autumn and spring. In addition, the high strategy was observed in smaller than 12 cm FL amount of large specimens might exploit all specimens’ particularly in summer and autumn and favorable animal materials in summer. herbivore dominant omnivore strategy was observed In Karamenderes River, feeding intensity of C. in larger than 18 cm FL specimens particularly in gibelio had a seasonal variation with high feeding summer and spring. The IRI % values are indicative
Partal and Yalçın Özdilek 2019 - LimnoFish 5(1): 6-16 15 of instant feeding, so animal materials are digested Bolnick DI, Amarasekare P, Araújo MS, Bürger R, Levine faster, especially in hot summer and autumn seasons JM, Novak M, Rudolf VHW, Schreiber SJ, Urban MC, (Windell 1978). Vasseur DA. 2011. Why intraspecific trait variation The increase in the trophic level with increasing matters in community ecology. Trends Ecol Evol. of the fish length was found in many studies as a 26(4):183-192. doi: 10.1016/j.tree.2011.01.009 natural process (Weber and Brown 2013). In this Cortés E. 1999. Standardized diet compositions and study, the finding that large specimens have high trophic levels of sharks. ICES J Mar Scı. trophic level in spring and summer is an anticipated 56(5):707–717. result. However, a decrease in the trophic level of doi: 10.1006/jmsc.1999.0489 larger than 20 cm in FL might be explained by food Costa MJ, Cabral HN. 1999. Changes in the Tagus nursery availability. 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LIMNOFISH-Journal of Limnology and Freshwater Fisheries Research 5(1): 17-26 (2019)
Turunçgil Kabuk Yağlarının Gökkuşağı Alabalığı (Oncorhynchus mykiss) Filetolarının Raf Ömrü Üzerine Etkileri
Pınar OĞUZHAN YILDIZ
Ardahan Üniversitesi, Mühendislik Fakültesi, 75000 Ardahan-Türkiye
ÖZ MAKALE BİLGİSİ
Bu çalışmada farklı konsantrasyonlardaki (%0,5 ve %1) turunçgil kabuk ARAŞTIRMA MAKALESİ yağlarının (limon, portakal ve bergamot) gökkuşağı alabalığı (Oncorhynchus mykiss) filetolarının raf ömrü üzerine etkileri araştırılmıştır. Örnekler kontrol Geliş : 14.05.2018 grubu (A), %0,5 limon uçucu yağı ilave edilmiş (B), %1 limon uçucu yağı ilave Düzeltme : 17.10.2018 edilmiş (C), %0,5 portakal uçucu yağı ilave edilmiş (D), %1 portakal uçucu yağı Kabul : 21.10.2018 ilave edilmiş (E), %0,5 bergamot uçucu yağı ilave edilmiş (F) ve %1 bergamot uçucu yağı ilave edilmiş (G) olmak üzere yedi gruba ayrılmıştır. Balık Yayım : 25.04.2019 örneklerinin mikrobiyolojik (toplam aerobik mezofilik bakteri, toplam psikrotrofik bakteri, laktik asit bakterileri, Enterobacteriaceae) kimyasal (pH, DOI:10.17216/LimnoFish.423440 toplam uçucu bazik azotu, thiobarbitürik asit reaktif maddeler) ve duyusal (renk, koku, lezzet, genel kabul edilebilirlik) özellikleri incelenmiştir. Depolama süresi SORUMLU YAZAR boyunca mikrobiyolojik ve kimyasal özellikler açısından kontrol grubu ile [email protected] uygulama grupları arasında önemli değişiklikler tespit edilmiştir (p<0,05). En Tel : +90 478 211 75 75 düşük ve en yüksek toplam aerobik mezofilik bakteri, toplam psikrotrofik bakteri, laktik asit bakteri ve Enterobacteriaceae sayıları sırasıyla 2,54-7,92, 2,69-8,03, 2,00-5,03 ve 2,00-4,09 olarak bulunmuştur. Çalışmadan elde edilen sonuçlara göre farklı konsantrasyonlarda turunçgil uçucu yağları ilavesinin bakteri sayısını önemli ölçüde azalttığı ve örneklerin bozulmasını geciktirdiği belirlenmiştir. Anahtar kelimeler: Gökkuşağı alabalığı, turunçgil kabuk yağları, raf ömrü, kalite özellikleri
Influences of Citrus Peel Oils on Shelf Life of Rainbow Trout (Oncorhynchus mykiss) Fillets Abstract: In this study, the effects of different concentrations (0.5% and 1%) citrus peel oils (lemon, orange and bergamot) on shelf life of rainbow trout (Oncorhynchus mykiss) fillets were researched. Samples were divided into seven groups: control (A), 0.5% [v/v] lemon EO added (B), 1% [v/v] lemon EO added (C), 0.5% [v/v] orange EO added (D), 1% [v/v] orange EO added (E), 0.5% [v/v] bergamot EO added (F) and 1% [v/v] bergamot EO added (G). Microbiological (total aerobic mesophilic bacteria, total psychrotrophic bacteria, lactic acid bacteria, Enterobacteriaceae), chemical (pH, thiobarbituric acid reactive substances and total volatile base nitrogen) and sensory (colour, odour, taste, general acceptance) properties of fish samples were investigated. During the storage period, significant changes were observed between the control group and treatment groups in terms of microbiological and chemical properties (p <0.05). The lowest and highest total aerobic mesophilic bacteria, total psychrotrophic bacteria, lactic acid bacteria and Enterobacteriaceae counts were found 2.54-7.92, 2.69-8.03, 2.00-5.03 and 2.00-4.09, respectively. According to the results obtained in this study, it was determined that the addition of citrus essential oils at different concentrations significantly reduced the number of bacteria and delayed the deterioration of the samples. Keywords: Rainbow trout, citrus peel oils, shelf life, quality properties
Alıntılama Oğuzhan Yıldız P. 2019. Turunçgil Kabuk Yağlarının Gökkuşağı Alabalığı (Oncorhynchus mykiss) Filetolarının Raf Ömrü Üzerine Etkileri. LimnoFish. 5(1): 17-26. doi: 10.17216/LimnoFish.423440
Giriş Dünyada ve Türkiye’de yetiştiriciliği en yoğun Beslenmede önemli bir yer tutan hayvansal ve yaygın olarak yapılan alabalık türü gökkuşağı kökenli proteinlerin temininde su ürünleri alabalığı (Oncorhynchus mykiss)’dır (Oğuzhan 2004; önemli kaynaklardan birisidir. Su ürünleri Sarıeyüpoğlu vd. 2017). Gökkuşağı alabalığı yüksek içerisinde de ilk sırayı balıklar almaktadır (Aras vd. adaptasyon ve yemden yararlanma yeteneği, suni 2000). yöntemlerle yumurta alımının kolaylığı, kuluçka
18 Oğuzhan Yıldız 2019 - LimnoFish 5(1): 17-26 sürelerinin kısalığı ve hastalıklara karşı dayanıklı yağı ilave edilmiş, D: %0,5 portakal uçucu yağı ilave olmasından dolayı tercih edilen bir türdür (Canyurt edilmiş, E: %1 portakal uçucu yağı ilave edilmiş, F: 1978, 1983; Emre ve Kürüm 1998; Oğuzhan 2004). %0,5 bergamot uçucu yağı ilave edilmiş ve G: %1 Gıdalarda sentetik ve doğal katkı maddeleri uzun bergamot uçucu yağı ilave edilmiş örneklerden yıllardan beri koruyucu olarak kullanılmaktadır. oluşmuştur. Ancak sentetik kökenli maddelerin sağlığa olan Turunçgil kabuk yağları bir fırça yardımıyla zararlı etkilerinden dolayı kullanımları filetoların her iki yüzeyine sürülmüştür. Filetonun sınırlandırılmış ve doğal katkı maddelerine olan önce bir yüzeyine, biraz kuruduktan sonra (5 dakika) yönelim her geçen gün artmıştır (Duman vd. 2012; ise diğer yüzeyine sürülmüştür. Daha sonra filetolar Kuş 2012; Mutlu ve Bilgin 2016). köpük tabaklar içine konularak streç filmle Su ürünlerinde doğal antioksidan kaynağı olarak kaplanmış ve buzdolabı (4°C±1) koşullarında çok sayıda bitki türü (biberiye, kekik, adaçayı, depolanmıştır. Filetolar depolamanın 0., 3., 6. ve 9. karanfil ve sarımsak) kullanılmıştır (Pazos vd. 2005; günlerinde mikrobiyolojik, kimyasal ve duyusal Serdaroğlu ve Felekoğlu 2005; Erkan 2012; özellikleri bakımdan incelenmiştir. Araştırma iki Çetinkaya 2013; Mutlu ve Bilgin 2016). tekerrürlü olarak yürütülmüştür. Narenciye yağları da antimikrobiyal ve Mikrobiyolojik Analizler antioksidan aktivitesi yüksek yağlardandır. Limon Mikrobiyolojik analizler petri yüzeyine yayma (Citrus limon), portakal (Citrus sinensis) ve metodu kullanarak hesaplanmıştır. Balık örneğinden bergamot (Citrus bergamia) narenciye familyasından 25 g tartılmış, sterilstomacher torbaya alınmış ve (Rutaceae) olup, esans yağ ve flavonoidler üzerine 225 ml steril serum fizyolojik (%0,85 NaCl, turunçgillerin kabuk kısmında yoğun bir şekilde Riedel-de Haën 13423) ilave edilerek stomacher bulunmaktadır. Bu kısımlar limonen ve linalool gibi cihazında (Lab Stomacher Blender 400-BA 7021 uçucu yağları oldukça yüksek miktarlarda Sewardmedical, England) 2 dakika homojenize içermektedirler. Narenciye yağları; gıda, kozmetik, edilmiştir. TAMB ve TPAB sayımları için Plate Count eczacılık, parfümeri ve kimya endüstrisi gibi birçok Agar (PCA, Oxoid CM0325) kullanılmıştır. TAMB alanda yaygın bir kullanım alanına sahiptir. Özellikle 30°C’de 2 gün, TPAB ise 7°C’de 7 gün inkübe de limon yağı gıda ve içecek endüstrisinde oldukça edilmiştir. LAB sayımı için Man Rogosa Sharpe Agar önemlidir. Limon ve bergamot yağı, antimikrobiyal (MRS, OxoidCM0361) kullanılmış ve ekimi yapılan ajan olarak başarıyla kullanılan uçucu yağlar arasında petri plakları 30°C’de 2 gün inkübe edilmiştir. yer almaktadır. Portakal kabuk yağının önemli Enterobacteriaceaesayımı için VRBD (Violet Red bileşeni olan limonenin antioksidan aktivitesinin Glucose) Agar (Oxoid CM0485) kullanılmış ve oldukça yüksek olduğu yapılan çalışmalarla da 30C’de anaerobik şartlarda 2 gün inkübe edilmiştir bildirilmiştir (Turhan vd. 2006; Min-Hsiung 2008; (Gokalp vd. 2001). Roberto vd. 2010; Sánchez-Gonzálezvd. 2011; Erhan ve Bölükbaşı Aktaş 2017). Kimyasal Analizler Bu çalışmada, farklı konsantrasyonlardaki (%0,5 TVB-N (Toplam uçucu baz azotu) Analizi ve %1) turunçgil kabuk yağlarının (limon, portakal Toplam Uçucu Bazik Azot (TVB-N) miktarı ve bergamot) gökkuşağı alabalığı (O. mykiss) Malle ve Tao (1987) tarafından yapılan yönteme göre filetolarının raf ömrü üzerine etkileri 9 günlük belirlenmiştir. Bu yöntemde 40 g balık örneğine 80 depolama süresi boyunca araştırılmıştır. ml %7,5’luk TCA çözeltisi ilave edilmiş ve Ultra- Turrax (IKA Werke T 25, Germany) ile 1 dakika Materyal ve Metot homojenize edilmiştir. Karışım 5 dakika santrifüj Araştırmada kullanılan gökkuşağı alabalıkları (400xg) edildikten sonra Whatman 3 (Whatman® Artvin İli Şavşat İlçesindeki özel bir işletmeden, Schleicher&Schuell CAT No:1001 125) filtre kağıdı turunçgil kabuk yağları ise (limon, portakal ve kullanılarak süzülmüştür. Elde edilen 25 ml filtrata 5 bergamot) Kardelen firmasından (Kardelen Tarım ml %10’luk NaOH (Riedel-de Haën 06203) ilave Ürünleri Ltd., Ankara) temin edilmiştir. Yaklaşık edildi. Daha sonra distilasyon cihazına (BEHR S1 200-250 g ağırlığında toplam 56 adet balık satın Steam Distillation Unit Labor-Technik GmbH, alınmıştır. Balıklar buz içerisinde strafor kutularda Dusseldorf/Germany) yerleştirilip erlen içerisine 10 soğuk zincire uygun olarak laboratuvara getirilmiştir. ml %4’lük borik asit çözeltisi (H3BO3 Merck Laboratuvar ortamına getirilen balıkların başları 1.00165.1000) ve 0,04 ml indikatör (0,1 g metil kesilmiş, iç organları uzaklaştırılmış ve derili kırmızısı Merck 1.06076.0025)+0,1 g brom kresol filetoları çıkarılmıştır. Elde edilen filetolar işleme yeşili (Merck 1.08121.0005)+100 ml etanol eklenmiş hazır hale getirilmiş ve A, B, C, D, E, F ve G olmak ve 50 ml distilat toplanıncaya kadar distilasyon üzere 7 gruba ayrılmıştır. A: kontrol grubu, B: %0,5 yapılmıştır. Elde edilen distilat 0,1 N H2SO4 çözeltisi limon uçucu yağı ilave edilmiş, C: %1 limon uçucu ile pembemsi renk oluşuncaya kadar titre edilmiş.
Oğuzhan Yıldız 2019 - LimnoFish 5(1): 17-26 19
TVB-N miktarı titrasyonda harcanan H2SO4 çözeltisi Chicago, IL,USA) paket programı kullanarak (n) dikkate alınarak aşağıdaki formülle göre varyans analizi yapılmış ve önemli bulunan hesaplanmıştır. varyasyon kaynaklarına ait ortalamalar Duncan çoklu karşılaştırma testi ile test edilmiştir. TVB-N (mg/100g)= n x 16,8 mg azot
TBARS (Thiobarbitürik asit reaktif maddeler) Bulgular Analizi Mikrobiyolojik Analiz Sonuçları TBARS değerinin belirlenmesi Lemon (1975) ve Farklı konsantrasyonlarda turunçgil kabuk Kılıç ve Richards (2003) tarafından verilen yönteme yağları (limon, portakal ve bergamot) ilave edilen göre yapılmıştır. Bu yöntemde 2 g örneğe 12 ml TCA gökkuşağı alabalığı filetolarına ait mikrobiyolojik çözeltisi %7,5 TCA (Trichloroacetic Acid, Riedel-de analiz bulguları Tablo 1’de verilmiştir. Haën 27242) %0,1 EDTA (Ethylenediaminetetra- Balık örneklerine ait en düşük TAMB sayısı acetic Acid, Riedel-de Haën 34549), %0,1 Propil (2,33±0,12 log kob/g) C örneğinde belirlenirken, en galat (Propyl Gallate, Fluka 48710-3 ml etanolde yüksek TAMB sayısı (7,92±0,19 log kob/g) A çözülür) ilave edilmiş ve 15-20 sn ultra-turrax’da örneğinde saptanmıştır. Depolama süresince gruplar homojenize edildikten sonra Whatman 1 arasında önemli farklılıklar olduğu belirlenmiştir (Whatman® Schleicher&Schuell CAT No:1001 125) (p<0,05). TMAB sayılarının turunçgil kabuk yağları filtre kâğıdı kullanılarak süzülmüştür. Süzüntüden 3 katkılı örneklerde kontrol grubuna daha düşük ml alınarak deney tüpüne aktarılmış, üzerine 3 ml olduğu görülmüştür. 0,02 M TBA (Thiobarbituric acid, Fluka 88481) Toplam psikrotrofik bakteri sayısı depolama çözeltisi eklenmiş ve karıştırılmıştır. Daha sonra günlerine ve gruplara göre önemli farklılıklar deney tüpleri 100ºC’de 40 dakika su banyosunda göstermiştir (p<0,05). En yüksek psikrotrofik bakteri bekletilerek, 5 dakika soğuk su içerisinde sayısı depolamanın son gününde A (8,03±0,02 log soğutulmuştur. 2000 g’de 5 dakika santrifüj (Hermle kob/g) örneğinde bulunurken, en düşük bakteri sayısı ZK 380, Germany) edildikten sonra depolamanın 0. gününde C (2,69±0,11 log kob/g) spektrofotometrede (Shimadzu Corporation, Japan) örneğinde tespit edilmiştir. 530 nm dalga boyunda absorbans okunmuştur. Depolama süresince en yüksek laktik asit bakteri Standardın hazırlanmasında TEP (1,1,3,3- sayısı kontrol grubu örneklerinde (9. gün 5,03±0,12 tetraetoksipropan) kullanılmış ve k (standart) değeri log kob/g), en düşük bakteri sayısı ise turunçgil 0,06 olarak hesaplanmıştır. Sonuç µmol malonaldehit kabuk yağları katkılı gruplarda (0. gün 2,00±0,00 log (MA)/kg olarak verilmiştir. kob/g)tespit edilmiştir (p<0,05). Depolama süresi TBARS= ((absorbans/ k (0,006) x 2/ 1000 x 6,8) boyunca tüm gruplarda LAB sayıları artmıştır. x 1000/örnek ağırlığı Enterobacteriaceae sayısı tüm gruplarda günlere göre artış gösterirken, en yüksek değer depolamanın pH Analizi son gününde A (4,09±0,14 log kob/g) örneğinde 10 g balık eti örneğine 100 ml saf su eklenmiş ve saptanmıştır. Muhafaza süresince gruplar arasında 1 dakika ultra-turrax’da (IKA Werke T 25, Germany) önemli farklılıklar olduğu tespit edilmiştir (p<0,05). homojenize edilmiştir. Daha sonra pH değerleri pH- metre (Schott Labstar pH, Germany) ile Gökalp vd Kimyasal Analiz Sonuçları (2001) tarafından belirlenen yönteme göre Farklı konsantrasyonlarda turunçgil kabuk ölçülmüştür. yağları (limon, portakal ve bergamot) ilave edilen gökkuşağı alabalığı filetolarına ait kimyasal analiz Duyusal Analiz bulguları Şekil 1’de verilmiştir. Duyusal analiz Kurtcan ve Gönül (1987) TVB-N değerlerinde depolama süresince önemli yöntemine göre yapılmıştır. Duyusal analiz için 5 artışlar saptanmıştır (p<0,05). En fazla artış kontrol kişilik panelist grubu oluşturulmuştur. Balık grubunda görülmüştür. Kontrol grubu örneklerde örnekleri yaklaşık 5 dakika mikrodalga fırında depolamanın 0. gününde TVB-N değeri 16,42 mg/100 pişirildikten sonra panelistlere sunulmuş ve g olarak bulunurken, depolama sonunda 26,24 panelistler örnekleri renk, koku, lezzet ve genel kabul mg/100 g’a ulaşmıştır. TVB-N değerinin turunçgil edilebilirlik açısından 1 ile 5 arasında kabuk yağları katkılı örneklerde daha düşük olduğu değerlendirmiştir. Puanlamada; 5 çok iyi, 4 iyi, 3 belirlenmiştir. En düşük TVB-N değeri C grubunda normal, 2 kötü ve 1 çok kötü olarak 10,33 mg/100 g olarak tespit edilmiştir. değerlendirilmiştir. En yüksek TBARS değeri (5,53 µmol MA/kg) İstatiksel Analiz kontrol grubu (A) örneklerinde, en düşük TBARS Araştırmadan elde edilen verilerin istatistiksel değeri (1,21 µmol MA/kg) ise %1 limon uçucu yağı olarak değerlendirilmesinde SPSS 20.00 (SPSS, Inc., ilave edilmiş C grubunda belirlenmiştir. Muhafaza
20 Oğuzhan Yıldız 2019 - LimnoFish 5(1): 17-26 süresince bütün gruplarda zamana bağlı olarak artış Depolama sonunda (9. gün) kontrol grubu örneklerde görülmüştür. Muhafaza süresi boyunca TBARS pH değerlerinde azalma görülmüştür. En düşük pH değeri bakımında gruplar arasında önemli farklılıklar değeri C grubunda 5,92 olarak bulunmuştur. (p<0,05) saptanmıştır. Muhafaza süresi boyunca pH değeri bakımında En yüksek pH değeri kontrol grubu örneklerde gruplar arasında önemli farklılıklar (p<0,05) tespit depolamanın 0. gününde 6,30 olarak ölçülmüştür. edilmiştir. Çizelge 1. Farklı konsantrasyonlarda turunçgil kabuk yağları (limon, portakal ve bergamot) ilave edilen gökkuşağı alabalığı örneklerinin mikrobiyolojik analiz sonuçları (log kob/g) Örnek Muhafaza Toplam Aerobik Toplam Psikrotrofik Enterobacteriaceae Günleri Mezofilik Bakteri Sayısı BakteriSayısı Laktik Bakter Sayısı Sayısı 0 4,07±0,03a 4,36±0,06a 2,16±0,06a 2,00±0,00a A 3 5,51±0,05b 5.86±0,05b 2,92±0,09b 2,97±0,04b 6 6,91±0,08c 7,02±0,02c 3,31±0,14c 3,55±0,13c 9 7,92±0,09d 8,03±0,02d 5,03±0,12d 4,09±0,14d 0 2,54±0,05a 2,88±0,02a 2,00±0,00a 2,00±0,00a B 3 3,05±0,07b 3,27±0,04b 2,25±0,04bc 2,25±0,04b 6 3.77±0,04c 3,85±0,09c 2,21±0,10b 2,66±0,02c 9 4.60±0,14d 4,99±0,07d 2,40±0,02d 2,81±0,06d 0 2,33±0,02a 2,69±0,11a 2,00±0,00a 2,00±0,00a C 3 2,96±0,05b 3,12±0,04b 2,11±0,04a 2,10±0,08a 6 2,35±0,07c 3,62±0,04c 2,07±0,09a 2,52±0,07b 9 4,12±0,04d 4,28±0,02d 2,66±0,14b 2,76±0,07d 0 2,74±0,04a 3,11±0,03a 2,00±0,00a 2,00±0,00a D 3 3,26±0,09b 3,70±0,05b 2,66±0,07b 2,48±0,09b 6 4,13±0,16c 4,03±0,06c 3,02±0,04c 2,82±0,07c 9 4,88±0,04d 5,29±0,07d 3,30±0,03d 2,95±0,05c 0 2,58±0,28a 3,01±0,03a 2,00±0,00a 2,00±0,00a E 3 3,13±0,07b 3,37±0,06b 2,53±0,07b 2,35±0,05a 6 3,95±0,04c 3,90±0,02c 2,81±0,14c 2,76±0,02c 9 4,73±0,08d 5,10±0,02d 3,07±0,05d 2,85±0,05c 0 3,02±0,04a 3,51±0,06a 2,00±0,00a 2,00±0,00a F 3 3,63±0,07b 4,02±0,04b 3,07±0,04b 2,92±0,05b 6 4,39±0,06c 4,51±0,10c 3,37±0,05c 3,11±0,04c 9 5,15±0,06d 5,68±0,03d 3,72±0,08d 3,36±0,04d 0 2,91±0,04a 3,31±0,04a 2,00±0,00a 2,00±0.00a G 3 3,40±0,01b 3,92±0,06b 2,92±0,06b 2,66±0,07b 6 4,20±0,08c 4,33±0,07c 3,21±0,08c 3,01±0,04c 9 4,98±0,05d 5,60±0,06d 3,49±0,06d 3,11±0,04c *Aynı sütundaki farklı harfler, Duncan çoklu karşılaştırma testine göre ortalamalar arasındaki önemli düzeydeki farklılıkları göstermektedir (p<0,05). Kısaltmalar; A: kontrol, B: %0,5 limon uçucu yağ katkılı örnek; C: %1 limon uçucu yağ katkılı örnek, D: %0,5 portakal uçucu yağ katkılı örnek, E: %1 portakal uçucu yağ katkılı örnek, F: %0,5 bergamot uçucu yağ katkılı örnek, G: %1 bergamot uçucu yağ katkılı örnek.
Oğuzhan Yıldız 2019 - LimnoFish 5(1): 17-26 21
a
b
c
Şekil 1. Farklı konsantrasyonlarda turunçgil kabuk yağları (limon, portakal ve bergamot) ilave edilen gökkuşağı alabalığı örneklerinin kimyasal analiz sonuçları A: kontrol, B: %0,5 limon uçucu yağ katkılı örnek; C: %1 limon uçucu yağ katkılı örnek, D: %0,5 portakal uçucu yağ katkılı örnek, E: %1 portakal uçucu yağ katkılı örnek, F: %0,5 bergamot uçucu yağ katkılı örnek, G: %1 bergamot uçucu yağ katkılı örnek. Farklı harfler, Duncan çoklu karşılaştırma testine göre ortalamalar arasındaki önemli düzeydeki farklılıkları göstermektedir (p<0,05).
22 Oğuzhan Yıldız 2019 - LimnoFish 5(1): 17-26
a b
c d
Şekil 2. Farklı konsantrasyonlarda turunçgil kabuk yağları (limon, portakal ve bergamot) ilave edilen gökkuşağı alabalığı örneklerinin duyusal analiz sonuçları A: kontrol, B: %0,5 limon uçucu yağ katkılı örnek; C: %1 limon uçucu yağ katkılı örnek, D: %0,5 portakal uçucu yağ katkılı örnek, E: %1 portakal uçucu yağ katkılı örnek, F: %0,5 bergamot uçucu yağ katkılı örnek, G: %1 bergamot uçucu yağ katkılı örnek.
Tartışma ve Sonuç log kob/g) ulaştığı saptanmıştır. Can vd. (2011), Bu çalışmayla farklı konsantrasyonlarda (%0,5 aynalı sazan (Cyprinus carpio carpio L., 1758) ve %1) turunçgil kabuk yağları (limon, portakal ve balıklarını Öjenol ile farklı sürelerde salamura bergamot) ilave edilen gökkuşağı alabalığı yaparak, 4°C’ de 9 gün süreyle depolamışlar ve A filetolarının buzdolabı (4°C±1) şartlarındaki raf ömrü grubu (6 saat) örneklerinde depolamanın 9. gününde araştırılmıştır. TMAB sayısı 6,19 log kob/g iken, B (12 saat) ve C (24 saat) grubu örneklerinde sırasıyla 3,03 ve 2,13 log Mikrobiyolojik Değişimler kob/g olarak bulmuşlardır. Bu çalışma sonucu Gıdaların mikrobiyolojik yükü, gıdanın kalitesi mevcut çalışmayla farklılıklar göstermektedir. Bu ile doğrudan ilişkilidir. Su ürünlerinde raf ömrünün durumun uygulanan esansiyel yağ ve balık türünün belirlenmesinde mikrobiyolojik analizler en sık farklı olmasından dolayı kaynaklandığı kullanılan yöntemlerden biridir (Mutlu ve Bilgin düşünülmektedir. Duman vd. (2012) biberiye ve 2016). 9 günlük depolama süresi boyunca örneklere kekik uçucu yağlarının kerevitlerin (Astacus TMAB, psikrotrofik, Enterobacteriaceae ve LAB leptodactylus Esch., 1823) raf ömrü üzerine analizleri yapılmıştır. Mikrobiyolojik analiz mikrobiyolojik etkisinin araştırıldığı çalışmalarında, sonuçlarına göre en düşük bakteri sayısı %1 limon kontrol grubuna oranla mikrobiyal yükün biberiye ve uçucu yağı ilave edilmiş C grubu örneklerinde tespit kekik esansiyel yağı katkılı örneklerde daha düşük edilmiştir. Depolama süresi boyunca örneklerin olduğunu belirtmişlerdir. Bu çalışma sonuçları bakteri sayıları artış göstererek, bu artışın istatiksel yapılan bu çalışma ile benzerlik göstermiştir. olarak önemli olduğu (p<0,05) bulunmuştur. Psikrotrofik bakteri sayısı kontrol grubunda Taze balıklarda bulunmasına müsaade edilen başlangıçta 4,36 log kob/g olarak saptanmıştır. Bu toplam bakteri sayısı değeri 7 log kob/g’dır (ICMSF değer muhafazanın 6. gününde yükselerek 1986). Bu değere A grubu örnekler muhafazanın 9. tüketilemez (7,02 log kob/g) değerine ulaşmış, diğer gününde ulaşırken, diğer gruplar depolama periyodu gruplar depolama periyodu sonunda bu değerin sonunda bu değerin altında bulunmuştur (Tablo1). altında belirlenmiştir (Tablo 1). Özyılmaz (2007), Toplam aerobik bakteri sayısının kontrol grubunda farklı dozlarda (5, 20, and 35 µl) püskürtme tekniği depolamanın 9. gününde tüketilemez değere (7,92 ile kekik esansiyel yağı uygulamasının gökkuşağı
Oğuzhan Yıldız 2019 - LimnoFish 5(1): 17-26 23 alabalığı (O. mykiss, Walbaum, 1972) filetolarının artış kontrol grubunda belirlenmiştir. TVB-N psikrofil bakteri faaliyetlerini önemli ölçüde değerlerinin turunçgil kabuk yağları katkılı yavaşlattığını belirtmiştir. Emir Çoban vd. (2018) örneklerde daha düşük olduğu görülmüştür. Benzer dondurulmuş gökkuşağı alabalığı (O. mykiss) sonuçlar Can vd. (2011) tarafından da bulunmuştur. filetoları üzerinde yaptıkları bir araştırmada Özpolat vd. (2017), farklı dozlardaki biberiye psikrofilik bakteri sayılarının karanfil yağı (%0,5 ve esansiyel yağlarının gökkuşağı alabalığı üzerine %1v/w) katkılı örneklerde, kontrol grubuna göre antimikrobiyel ve antioksidan etkilerini araştırdıkları daha düşük olduğunu tespit etmişlerdir. Bu çalışma çalışmalarında, TVB-N değerindeki yükselişin en sonuçları mevcut çalışma ile benzerlik göstermiştir. fazla kontrol grubunda, en az yükselişin ise %2 Kontrol grubunda LAB depolamanın 0. gününde oranında biberiye uygulanan grupta olduğunu ve tüm 2,16 log kob/g ve depolama sonunda ise 5,03 log gruplarda muhafaza süresi ile birlikte TVB-N kob/g olarak saptanmıştır. Muhafaza süresince değerinin istatistiki açıdan önemli derecede örneklerin laktik asit bakteri sayıları artış göstererek, yükseldiğini belirlemişlerdir (p<0,05). Güran vd. bu artışın istatiksel olarak önemli olduğu (p<0,05) (2015) kekik, karanfil ve biberiye yağları uygulanmış belirlenmiştir. LAB sayılarının turunçgil kabuk palamut (Sarda sarda Bloch, 1793) balıkları ile yağları katkılı örneklerde daha düşük olduğu yaptıkları çalışmalarında TVB-N değerlerinin kontrol görülmüştür. Duman vd. (2012) laktik asit bakterileri grubunda, diğer örneklere kıyasla daha yüksek üzerinde biberiye ve kekik esansiyel yağlarının etkili olduğunu rapor etmişlerdir. olduğunu bildirmişlerdir. Patır vd. (2015) farklı TBARS değeri, lipit oksidasyonunun derecesinin oranlarda asetik asit ile (%2 ve %4) hazırlanan belirlenmesinde yaygın olarak kullanılan önemli bir alabalık (O. mykiss) marinatları üzerine %0,1 ve kalite kriteridir (Günlü 2007; Oğuzhan 2011). %0,5’lik eugenolün etkisinin araştırıldığı çalışmada Muhafaza süresine bağlı olarak kontrol grubu ve laktik asit bakteri sayılarının Eugenol katkılı esansiyel yağ uygulanan gruplarda TBARS örneklerde, kontrol grubuna kıyasla daha düşük değerlerinde artışlar tespit edilmiştir. Depolama olduğunu saptamışlardır. Mevcut bu çalışma periyodu boyunca en yüksek TBARS değeri kontrol sonuçları araştırmacıların bulgularıyla benzerlik grubu örneklerinde belirlenirken, en düşük %1 limon göstermiştir. uçucu yağı ilave edilmiş C grubu örneklerinde Depolama süresince en yüksek saptanmıştır (Şekil 1b). Fernandez-Lopez vd. (2005), Enterobacteriaceae bakteri sayısı kontrol grubu (A) TBA değerlerinin biberiye, limon ve portakal yağları örneklerinde, en düşük bakteri sayısı ise %1 limon katkılı köftelerde kontrol grubuna kıyasla daha düşük uçucu yağı ilave edilmiş C grubunda bulunmuştur. olduğunu ve esansiyel yağların muhafaza süresine de Muhafaza süresi boyunca tüm gruplarda önemli etkilerinin (p<0,05) olduğunu Enterobacteriaceae sayıları artış göstermiş, ancak en vurgulamışlardır. Emir Çoban ve Can (2013) fazla artış kontrol grubunda tespit edilmiştir. gökkuşağı alabalıkları (O. mykiss) ile yürüttükleri bir Enterobacteriaceae bakteri sayılarının turunçgil çalışmalarında farklı konsantrasyonlarda (%0,5 ve kabuk yağları katkılı örneklerde daha düşük olduğu %1) biberiye esansiyel yağını kullanmışlar ve TBA belirlenmiştir. Mutlu ve Bilgin (2016) sıcak değerlerinin kontrol grubunda, biberiye yağı ilaveli dumanlanmış gökkuşağı alabalığı (O. mykiss) ile örneklere kıyasla daha yüksek olduğunu yapmış oldukları çalışmalarında zeytin yaprağı ve vurgulamışlardır. Her iki çalışmadaki sonuçlar, bu yağ gülü ekstraktlarının Enterobacteriaceae sayısı çalışmadan elde edilen değerlerle benzerlik üzerine etkili olduğunu bildirmişlerdir. Aynı şekilde göstermiştir. Duman vd. (2012) biberiye ve kekik Tassou (1996), limon ve kekik esansiyel yağlarını uçucu yağları uygulanmış kerevitler (A. leptodactylus uygulanmış çipura (Sparus aurata) filetolarında Esch., 1823) ile yaptıkları çalışmalarında, en yüksek enterobakteri sayısının kontrol grubuna kıyasla daha TBA değerinin kontrol grubunda, en düşük ise kekik düşük olduğunu rapor etmişlerdir. esansiyel yağı uygulanmış örneklerde olduğunu vurgulamışlardır. Kimyasal Değişimler Balık etinde pH değerinin 6,0 ile 6,5 arasında Balık ve balık ürünlerinin tazeliğin değişkenlik gösterdiği ve bu değerin muhafaza belirlenmesinde çok fazla kullanılan kimyasal süresince bozulma ile birlikte yükseldiği ve parametrelerden birisi de TVB-N değeridir (Köse ve tüketilebilirlik sınır değeri olarak da 6,8-7,0 kabul Koral 2005; Oğuzhan 2011; Mutlu ve Bilgin 2016). edildiği çeşitli araştırıcılar tarafından bildirilmiştir. Depolama periyodu boyunca en yüksek TVB-N pH değeri her zaman kesin bir kriter olmayıp değeri kontrol grubu örneklerinde, en düşük ise %1 duyusal, kimyasal ve mikrobiyolojik testlerle de limon uçucu yağı ilave edilmiş C grubu örneklerinde desteklenmesi gerekmektedir (Baygar vd. 2004; İnal saptanmıştır (Şekil 1a). Depolama süresi boyunca 2007; Özpolat vd. 2017). Depolama süresi boyunca tüm gruplarda TVB-N değeri artış göstermiş, en fazla tüm grupların pH değerlerinin dalgalanmalar
24 Oğuzhan Yıldız 2019 - LimnoFish 5(1): 17-26 gösterdiği ve istatistiksel olarak pH değerleri Sonuç olarak; turunçgil kabuk yağlarının arasındaki farkın önemli olduğu bulunmuştur gökkuşağı alabalığı filetolarının kimyasal ve (p<0,05). Yerlikaya vd. (2005) yapmış oldukları mikrobiyolojik özellikleri üzerine olumlu etki yaptığı çalışmalarında hamsi köftelerin pH değerlerinin ve raf ömrünün arttırılmasında pozitif sonuçlar depolama süresince arttığını rapor etmişlerdir. verdiği tespit edilmiştir. Duman vd. (2012) kontrol örnekleri ile biberiye ekstraktı uygulanmış örnekler arasında pH değeri Kaynaklar bakımından istatistiksel olarak önemli farklılıklar Akçay S. 2012. Antimikrobiyal madde içeren yenilebilir olmadığını bildirmişlerdir. Alçiçek (2011) sıvı filmlerin dumanlanmış balığın kalitesine etkisi. tütsülenmiş gökkuşağı alabalığı üzerinde yürüttükleri [Yüksek Lisans Tezi]. İstanbul Üniversitesi. 85 s. Alçiçek Z. 2011. The effects of thyme (Thymus vulgaris l.) çalışmalarında kontrol grubu ile farklı oil concentration on liquid-smoked vacuum-packed konsantrasyonlarda kekik yağı uygulanmış gruplar rainbow trout (Oncorhynchus mykiss Walbaum, 1792) arasında pH değeri bakımından istatistiksel açıdan fillets during chilled storage. Food Chem. 128: 683- önemli farklılıklar olduğu saptanmıştır (p<0,05). 688. doi: 10.1016/j.foodchem.2011.03.087 Duyusal Değişimler Aras NM, Kocaman EM, Aras MS. 2000. Genel su Panelistler tarafından tüm kriterler (renk, koku, ürünleri ve kültür balıkçılığının temel esasları. lezzet ve genel kabul edilebilirlik) açısından en fazla Erzurum: Atatürk Üniversitesi Ziraat Fakültesi kontrol (A) grubu örnekleri beğenilirken, en az %1 Yayınları No: 216. Atatürk Üniversitesi Ziraat bergamot uçucu yağı ilave edilmiş G grubu Fakültesi Ofset Tesisi, 295 p. Baygar T, Özden Ö, Üçok D. 2004. Dondurma ve örneklerinin beğenildiği belirlenmiştir (Şekil 2). çözündürme işleminin balık kalitesi üzerine etkisi. Kullanılan ekstraktların balıkta alışılmadık bir lezzet, Turk J Vet Anim Sci. 28:173-178. koku vermesi ve dolayısıyla genel kabul edilebilirlik Can ÖP, Yalçın H, Arslan A. 2011. Farklı sürelerde açısından tercih etmedikleri düşünülmektedir. Tüm öjenol’lü salamura solüsyonunda bekletilen aynalı parametreler açısından gruplar arasındaki farkın sazan balığı (Cyprinus carpio carpio L., 1758) istatistiksel olarak önemli olduğu tespit edilmiştir filetolarının kalite kriterlerinin değerlendirilmesi. (p˂0,05). Çalışmaya paralel olarak Duman vd. Paper presented at: International Symposium on (2012) duyusal analiz sonuçlarına göre panelistlerin Fisheries and Aquatic Sciences (FABA); Trabzon, en çok kontrol grubu örneklerini beğenirken, en az Turkey. kekik ilaveli grubu beğendiklerini bildirmişlerdir. Canyurt MA. 1978. Alabalık üretim. İzmir: Ege Üniversitesi Ziraat Fakültesi Zootekni Derneği Yayını Mutlu ve Bilgin (2016) tarafından yapılan başka bir 66 p. çalışmada ise gül ekstraklı grubun en beğenilmeyen Canyurt MA.1983. Alabalık yetiştiriciliği. Diyarbakır: grup olduğu vurgulanmıştır. Emir Çoban vd. (2018) Dicle Üniversitesi Ziraat Fakültesi Yıllığı. 2:7-42. dondurulmuş gökkuşağı alabalığı (O. mykiss) Çetinkaya S. 2013. Vakum paketli pişirilen (Sous-Vide) filetoları ile yürütülen bir araştırmada karanfil yağı gökkuşağı alabalığı (Oncorhynchus mykiss Walbaum, uygulanmış örneklerin duyusal parametreler (koku, 1792)’nın soğuk depolanması sırasında kalite renk, tekstür) açısından kontrol grubuna kıyasla daha özelliklerine doğal antioksidanların etkisi [Doktora fazla beğenildiğini rapor etmişlerdir. Sıcak Tezi]. Süleyman Demirel Üniversitesi. 142 s. tütsülenmiş gökkuşağı alabalığı (O. mykiss) ile Duman M, Emir Çoban Ö, Özpolat E. 2012. Biberiye ve yürütülen bir çalışmada balıklar karanfil, sarımsak ve kekik esansiyel yağları katkısının marine edilmiş kerevitlerin (Astacus leptodactylus Esch., 1823) raf kekik uçucu yağları ilave edilerek buğday proteini ömrüne etkisinin belirlenmesi. Kafkas Univ Vet Fak gluteninden filmleri ile kaplanmış, vakum Derg. 18(5):745-751. paketlenmiş ve +2±2 °C’de depolanmıştır. Uçucu doi: 10.9775/kvfd.2012.5975 yağlar ilave edilerek kaplama uygulanmış örneklerin Emir Çoban Ö, Can ÖP. 2013. The effect of active kaplama uygulanmamış örneklere kıyasla duyusal packaging film containing rosemary extract on the (genel görünüş, koku, tat ve doku) açıdan daha çok quality of smoked rainbow trout (Oncorhynchus beğenildiğini bildirmiştir (Akçay 2012). Emir Çoban mykiss). J Aquat Food Prod T. 22(4):361–370. ve Tuna Keleştemur (2017) tarafından yapılan bir doi: 10.1080/10498850.2011.652766 çalışmada Zataria multiflora Boiss. uçucu yağının Emir Çoban Ö, Tuna Keleştemur G. 2017. Qualitative farklı konsantrasyonlarını (%0, %0,2 ve %0,4) improvement of catfish burger using Zataria multiflora Boiss. essential oil. J Food Meas Charact. kullanarak ürettikleri yayın balığı (Silurus glanis, 11(2): 530-537. Linnaeus, 1758) burgerlerinde duyusal kriterler doi:10.1007/s11694-016-9420-2 (koku, tat, tekstür, genel kabuledilebilirlik) Emir Çoban Ö, Özpolat E, Karaton Kuzgun N. 2018. The bakımından en yüksek puanı %0,4 uçucu yağ ilave effect of clove oil on frozen storage stability and edilerek üretilen burgerlerin aldığı rapor edilmiştir. quality of rainbow trout (Oncorhynchus mykiss). Ege
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Biberiye ürünlerinde kalite kontrolü ve laboratuvar uygulama esansiyel yağı ile dekontaminasyonun gökkuşağı kılavuzu. Erzurum: Atatürk Üniversitesi Ziraat alabalıklarının (Oncorhynchus mykiss, W. 1972) kalite Fakültesi Yayını 268 p. özellikleri üzerine etkisi. FÜ Sağ.Bil Vet Derg. Günlü A. 2007. Yetiştiriciliği yapılan deniz levreğinin 31(2):131-136. (Dicentrarchus labrax L. 1758) dumanlama sonrası Özyılmaz A.2007. Gökkusağı alabalığı (Oncorhynchus bazı besin bileşenlerindeki değişimler ve raf ömrünün mykiss, Walbaum, 1972) filetolarında kekik eterik belirlenmesi [Doktora Tezi]. Süleyman Demirel yağı kullanımının raf ömrü üzerine etkisi [Yüksek Üniversitesi. 123 s. Lisans Tezi]. Mustafa Kemal Üniversitesi. 57 s. Güran HS, Öksüztepe G, Emir Çoban Ö, Inicili GK. 2015. Patır B, Can ÖP, Emir Çoban Ö, Özpolat E. 2015. 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LIMNOFISH-Journal of Limnology and Freshwater Fisheries Research 5(1): 27-33 (2019)
Potential Distribution of the Amphibian Pathogen, Batrachochytrium dendrobatidis in the Eastern Black Sea Region of Turkey
Uğur Cengiz ERİŞMİŞ
Afyon Kocatepe University, Faculty of Sciences and Literatures, Molecular Biology & Genetics Department, Afyonkarahisar, Turkey
ABSTRACT ARTICLE INFO
Although Batrachochytrium dendrobatidis, pathogen for amphibians, has been RESEARCH ARTICLE reported in Anatolia, its geographical distribution, as well as its impact on the amphibians in Turkey, remained obscure. In this study, 62 adult individuals Received : 05.10.2018 belonging to ten different species (Pelodytes caucasicus, Rana dalmatina, Rana Revised : 10.12.2018 macrocnemis, Bufo bufo, Bufo verrucosissimus, Bufotes variabilis, Hyla savignyi, Accepted : 15.01.2018 Pelophylax ridibundus, Ommatotriton ophryticus, and Mertensiella caucasicus) were collected from five wetland habitats in Eastern Black Sea Region of Turkey. Published : 25.04.2019 The prevalence and the intensity of B. dendrobatidis infections in all the individuals were investigated by using quantitative Real-time-PCR technique and DOI:10.17216/LimnoFish.467527 the presence of B. dendrobatidis infection was reported for the first time in 13 of the 62 individuals collected from 10 amphibian species from Eastern Black Sea * CORRESPONDING AUTHOR Region of Turkey. The intensity of B. dendrobatidis infection ranged from [email protected] 403.520 to 534.280 genomic equivalents (GE) was detected. The highest GE Phone : +90 272 228 13 39 between amphibian species were determined in P. caucasicus (534.280 GE) in Uzungöl (Çaykara-Trabzon) and B. bufo (504.00 GE) in Lake Karagöl (Şavşat- Artvin). Keywords: Chytridiomycosis, Batrachochytrium dendrobatidis, Anatolia, Amphibia
Doğu Karadeniz Bölgesinde Amfibi Patojeni Batrachochytrium dendrobatidis’in Potansiyel Dağılımı Öz: Amfibi patojeni Batrachochytrium dendrobatidis Anadolu’da rapor edilmesine rağmen, hem coğrafik dağılımı hem de Türkiye amfibileri üzerindeki etkisi hala belirsizdir. Bu çalışmada, Doğu Karadeniz Bölgesindeki beş sulak alandan on farklı amfibi türlerine (Pelodytes caucasicus, Rana dalmatina, Rana macrocnemis, Bufo bufo, Bufo verrucosissimus, Bufotes variabilis, Hyla savignyi Pelophylax ridibundus, Ommatotriton ophryticus, Mertensiella caucasicus) ait 62 ergin birey toplandı. Kantitatif Real-time PCR tekniği ile tüm bireylerde B. dendrobatidis enfeksiyonlarının prevalansı ve yoğunluğu ile araştırıldı ve Doğu Karadeniz Bölgesi’ndeki 10 amfibi türünden toplanan 62 bireyin 13’ ünde B. dendrobatidis enfeksiyonu varlığı ilk kez rapor edildi. Enfeksiyon yoğunluğu 403,520-534,340 genomik eşdeğerler arasında değiştiği belirlendi. Amfibi türler arasında en yüksek genomik eşdeğer Uzungöl’ deki P. caucasicus (534,280) ve Karagöl'de B. bufo (504,00) saptandı. Anahtar kelimeler: Chytridiomycosis, Batrachochytrium dendrobatidis, Anadolu, Amphibia
How to Cite Erişmiş UC, 2019. Potential Distribution of the Amphibian Pathogen, Batrachochytrium dendrobatidis in the Eastern Black Sea Region of Turkey. LimnoFish. 5(1): 27-33. doi: 10.17216/LimnoFish.467527
Introduction such as host population density, habitat, and age Infectious diseases are one of the factors structure may influence the rate at which chytrid implicated in the declines and extinctions of spreads through the environment (Daszak et al. amphibians in worldwide. Batrachochytrium 1999). According to Ron (2005), Bd was predicted to dendrobatidis (Bd) is a fungus that colonizes spread in Anatolia, but the geographic distribution of amphibian skin and the associated disease, Bd and its effect on Turkish amphibians is poorly chytridiomycosis, can disturb cutaneous respiration understood (Farrer et al. 2011). Though 26 Turkish and osmoregulation and result in the death of the host amphibian species are listed in the International (Carver et al. 2010). Differences in ecological factors Union for Conservation of Nature (IUCN) Red List,
28 Erişmiş 2019 - LimnoFish 5(1): 27-33 investigation of the decrease in frog population has Hence, the location of pathogens and susceptible to become mandatory (Başkale et al. 2013). Only two species are needed to be determined in Turkey. works were carried out on the distribution of Bd in Management strategies for the containment of Bd Turkey. Previously, Göçmen et al. (2013) reported spreading include the detection of wild and captive that one of two P. bedriagae specimens from Göynük populations infected with chytrid disease, the Canyon (Antalya) was found as positive for Bd. identification of infected geographical areas, and the Erişmiş et al. (2014) reported Bd infecting wild P. control of infected animal’s movement from one ridibundus, H. orientalis, B. variabilis as well as location to another. endemic Beysehir frogs P. caralitanus in West Therefore, the main objective of the present study Anatolian Region and the District Lakes of South was to determine Bd infected amphibian species Western Turkey. through quantitative polymerase chain reactions Thirty-six amphibian species were recorded in (qPCR) (Kriger et al. 2006; Hyatt et al. 2007) in the Turkey. Due to Turkey’s geographical position, Eastern Black Sea Region of Turkey. different species spread in different regions and they are exposed to a great number of threats (Şekercioğlu Materials and Methods et al. 2011). This includes a number of restricted and The study was carried out in 6 different rare amphibian species such as Rana tavasensis areas [Uzungöl (Çaykara-Trabzon, UZL), Karagöl (Franzen et al. 2008), R. holtzi (Yildiz and Göçmen (Şavşat-Artvin, KRL), Sahara Natural Park (Şavsat- 2012), R. macrocnemis (Veith et al. 2003), P. Artvin, SNP), Ardeşen (Rize, ARD), Lake Şavsat caralitanus (Bülbül et al. 2011). If such species were (Şavşat-Artvin, SVT)] at 39 to1876 m elevation (E) susceptible to the fungal infection, the local and in the eastern Black Sea region (EBS) of Turkey isolated populations might easily become extinct. (Figure 1).
Figure 1. Map showing the collecting sites. The specimens were collected during the summer studied in collected 10 species [P. caucasicus (Pc), season of 2014 (June through August). Air R. dalmatina (Rd), R. macrocnemis (Rm), B. bufo temperature, water pH, and humidity (H) were (Bb), B. verrucosissimus (Bv), B. variabilis (Bvs), H. recorded during the fieldwork (Table 1). For each savignyi (Hs) P. ridibundus (Pr), O. ophryticus (Oo), captured animal, surveyors recorded its GPS M. caucasicus (Mc)]. coordinates. To prevent the transfer of infected To determine whether the animal was infected materials among sites, we rinsed all equipment with with Bd or not by PCR analysis, tissue samples were 5% bleach before entering each location. All of the collected using swab method with a sterile cotton tip frogs were handled with latex gloves and gloves were swab to take the keratinized tissues where Bd discarded after examination of each animal (Bai et al. zoospores were highly concentrated (Marantelli et al. 2010). The distribution of zoosporangia of Bd was 2004). During Bd sampling process, each individual
Erişmiş 2019 - LimnoFish 5(1): 27-33 29 was swabbed 30 times. We followed the standardized compared among within sites or the species due sampling protocol detailed by Hyatt et al. (2007). to the low statistical power of small sample Samples were stored in 95% ethanol and were kept sizes for each species at a site. In the localities with on ambient temperature (≥10°C) in the field and positive Bd, we used the zonal statistic transported back to the laboratory and stored in a −80 routine to extract from the digital maps the °C freezer (Hyatt et al. 2007). The intensity of environmental variables values from each point infection in all samples was determined by using (ArcView 3.2, Spatial Analyst). These values were qPCR (Boyle et al. 2004; Hyatt et al. 2007), with the also used to run Principal Component modifications of methods described by Boyle et al. Analysis (PCA; implemented in XLSTAT v.3.0) to (2004). It was extracted nucleic acids using 50 μl visualize the degree of clustering in environmental PrepMan Ultra (Applied Biosystems), and the tip of space among EBS region of Turkey where Bd was the swab was used instead of a toe. To ensure the found. integrity of our results, a negative control (dH2O) was run in triplicate on every 96-well PCR plate (Kriger Results et al. 2006). We constructed a standard curve to We swabbed 62 individuals from the 8 genera, determine the zoospore load. A standard curve was including 10 species that occur in EBS region of constructed from the control reactions containing Turkey. The prevalence and the intensity of Bd 100, 10, 1 and 0.1 Bd zoospores and the concentration infections in all the individuals were investigated by determined for the test samples expressed as the using quantitative real-time-PCR technique and the number of zoospore equivalents. The intensity of presence of Bd infection was reported for the first infection was measured as the number of genome time in 13 of the 62 individuals collected from 10 equivalents (GE) per swab, calculated by multiplying amphibian species from EBS Region of Turkey the GE values generated during the qPCR by the (Figure 2, Table1). dilution factor of the template DNA. Swabs were Bd was not detected only in ARD region. We determined the presence of Bd infection in 13 out of categorized as Bd positive at ≥1 GE and as Bd- 62 (20.9%) samples comprising six species: negative at <1 GE (Kriger et al. 2006; Hyatt et al. Pelobates caucasicus (Caucasian type-specific), 2007; Anna et al. 2011; Erismis et al. 2014). All Bufo bufo, B. verrucosissimus, Bufotes variabilis, analyses were performed in triplicate. The Pelophylax. ridibundus, Ommatotriton ophryticus percentages of infected individuals and GE were not (Table 1).
Figure 2. Map of Bd prevalence on EBS of Turkey. EBS from all states sampled tested positive and negative for Bd, Positive (black) and negative (white) proportions by the state were indicated by pie charts.
30 Erişmiş 2019 - LimnoFish 5(1): 27-33
Table 1. Prevalence (Prev) and Average Genomic Equivalents (GE) of Bd in Eastern Black Sea Region of Turkey
Air H E N Prev. Species Locality Coordinates GEs ˚C % (a.s.l) (+ve) (%)
Pc UZL 40˚3723N - 40˚1650E 26.0 56.0 1164 12(2) 16.0 527±16
Rd UZL 40˚3624N - 40˚1848E 19.5 78.0 1271 1(0) - - Rm. UZL 40˚3518N - 40˚2119E 18.5 82.0 1701 2(0) - - UZL 40˚3707N - 40˚1845E 19.5 78.0 1272 4(0) - - Bb KRL 40˚5620N - 43˚2330E 27.0 65.0 1600 4(1) 25.0 504.00 Bv UZL 40˚3711N - 40˚1736E 19.5 78.0 1278 10(1) 10.0 436.00 Mc UZL 40˚3527N - 40˚2045E 27.5 72.3 1702 3(0) - - Bvs SNP 41˚1424N - 41˚1312E 23.0 77.0 1876 2(1) 0.50 463.00 Hs ARD 41˚1113N - 40˚5919E 28.0 68.0 39.0 2(0) - - SNP 41˚1424N - 41˚1312E 23.0 77.0 1876 15(5) 33.3 490.00 Pr KRL 41˚1832N - 42˚2857E 26.0 65.0 1600 4(1) 25.0 480.00 Oo SVT 41˚1747N - 42˚2837E 30.5 66.0 1409 3(2) 66.6 405±9.0 Total 62(13) 20.9 473.66±18.44 Sampling localities and examined frog species abbreviations used; UZL: Uzungöl/Trabzon, KRL: Karagöl/Artvin, SNP: Sahara Natural Park/Artvin, ARD: Ardeşen/Artvin, SVT: Savsat Lake/Artvin; Pc: Pelodytes caucasicus, Rd: Rana dalmatina, Rm: Rana macrocnemis, Bb: B. bufo, Bv: Bufo verrucosissimus, Mc: Mertensiella caucasicus; Bv: B. variabilis, Hs: Hyla savignyi, Pr: Pelophylax ridibundus, Oo: O. ophryticus respectively, and H: Humidity, E: Elevation. GE genomic equivalent (including positive samples and negative samples, GE represents the burden of infection with Bd). Bd was detected at 4 of 5 sites in EBS Region of However, the average GEs among the four regions Turkey were not being specifying Bd may be due to (UZL, KRL, SNP, SVT) individuals of frogs infected the small number of samples (Figure 1). B. bufo were by Bd were analyzed through multiple comparisons sampled at two locations (UZL and KRL), only one of based on a Tukey-HSD post-hoc test, which indicated these locations tested positive for Bd. Although P. a not significant difference among them (F = 3.27, df ridibundus were sampled at two regions (SNP and = 3;9, p = 0.08). Increasing suitability for both KRL) but 6 tested positive for Bd. The population of prevalence and GE of Bd was widely distributed on both P. ridibundus and B. bufo (at KRL) with a EBS regions of Turkey but was detected lowest in the prevalence of 25%. However, we detected Bd in only ARD region (Tablo 1). one specimen of B. verrucosissimus (n=10). Final map resulted with areas highly suitable for Furthermore, we did not detect any Bd on R. the presence of Bd (Niche Overlap Index (NOI) > dalmatina, R. macrocnemis, B. bufo, and M. 0.70) dispersed irregularly overall EBS Region of caucasicus (at UZL), H. savignyi (at ARD). In Turkey (Figure 3). There are localities with highly addition, Northern banded newts (O. ophryticus) suitable for the fungus in north regions of were notable for their highest Bd infection rate at SVT phytogeographic provinces of Trabzon and Artvin with the prevalence of 66.6 % than other frog species (Northeast of Turkey). NOI was varied among (Table 1). Therefore, the prevalence of Bd infection regions and was represented by different percentages on the populations of 10 frogs species among EBS of covered surface. Areas with 0.70 > OI < 1 (highest regions (UZL, KRL, SNP, SVT) did differ suitability for chytrid development) covered only significantly (χ2 = 8.43, df = 3, P0.05> 0.03). 32.72 % of the total surface while areas with 0.50 > We also detected the rate of Bd infection as the OI < 0.70 covered 56.86 %. Areas with 0.00 > OI < mean number of GE per sample in 3 replicates. The 0.50 only 10.42 of the total surface of EBS region of mean number of GE for individual positive samples Turkey (Figure 3). ranged from 405±9.0 (for O. ophryticus at SVT With eigenvalues >1, Principal Component I was region) to 527±16.0 (for P. caucasicus at UZL positively correlated with (1) Elevation, (2) mean region). The highest intensity of zoospores was found annual temperature, (2) precipitation of wettest at P. caucasicus (527±16.00) at UZL region followed explaining 89.6% of the variance of the system. by B. bufo (504.00) at KRL region (Table 1). Principal Component II explained 29.0% and was
Erişmiş 2019 - LimnoFish 5(1): 27-33 31 highly positively correlated only with seasonal temperature ranged from 5 ̊C (UZL and SVT) to temperature. The environmental variables in the 23.5 ̊C (SNP and SVT), moreover in areas where the localities where Bd. was found show that suitable OI = 0.89. In addition, our analysis shows that the locations for the fungus are possible across a wide presence of the fungus in EBS region of Turkey is range of habitats (Table 1). In the localities with related to precipitations between 68 mm (UZL and known presence of the fungus, the annual mean SVT) and 235 mm (UZL and SVT).
Figure 3. Niche Overlap Index (NOI) map of the potential distribution of Bd in EBS region of Turkey Discussion variabilis, H. savignyi and M. caucasicus. In Distribution models showed that Bd has likely addition, Northern banded newts (O. ophryticus) already spread to most climatically suitable regions were notable for their higher Bd infection rate at SVT (Fisher et al. 2009). Turkey's diverse regions have prevalence of 75% than other frogs species. P. different climates because of irregular topography. ridibundus were sampled at two locations (SNP and Black Sea region has an oceanic climate (Köppen KRL) but 6 tested positive for Bd. We found a climate classification: Cfb), wet and humid (summer moderate Bd rate (20.09 %, N = 62) and low zoospore 23°C, winter 7°C) (Sensoy et al. 2008). Our previous loads (473.66±18.44). The number of zoospores study suggests that the Anatolian climate is indeed increases during infection. Low numbers may favorable for the spread of chytridiomycosis (Erismis represent an earlier stage of infection, but the et al. 2014). Intercalarily, Bd has been found in five shedding of the skin may also contribute to low mainlands including North and South America, counts. We observed a widespread prevalence of Bd Europe, Oceana, Africa, and Asia. However, there zoospore in apparently healthy adult amphibians in have only been two published studies up to date the study areas. Recent work suggests that Bd may describing the presence of Bd in Anatolia at the produce tiny, non-pathogenic resting spores that crossroads of three continents. Therefore, it is not attach to the amphibian skin surface but without surprising that the Bd present in Anatolia. causing disease (Di Rosa et al. 2007). The competing In this study, a total of 62 specimens from 8 hypothesis contends that chytrid is endemic to many genera, including 10 species was sampled. We regions and that climate or other factors have altered detected the presence of Bd infection in 13 of 62 the host-pathogen relationship, resulting in the recent (20.09%) samples comprising four species: P. outbreaks of chytridiomycosis (Morehouse et al. caucasicus (Caucasian type-specific), B. bufo, B. 2003; Weldon et al. 2004). As is also known adults verrucosissimus, P. ridibundus, O. ophryticus (Table may be protected by acquired immunity (Richmond 1). Bd was detected at 4 of 5 sites. We detected Bd in et al. 2009) and thus may clear or prevent infections only one from ten swamp samples of B. more efficiently than juveniles that are naive to Bd, verrucosissimus. We did not detect any Bd on R. infects some amphibian species with little negative dalmatina, R. macrocnemis, B. bufo (at UZL), B. effects on the host and do not die therefore may serve
32 Erişmiş 2019 - LimnoFish 5(1): 27-33 as reservoirs of the disease (Mazzoni et al. 2003). Bai C, Garner TWJ, Garner, LiY. 2010. First evidence of Many amphibian species such as Xenopus spp, R. Batrachochytrium dendrobatidis in China: discovery catesbeiana, and B. marinus carry this disease, also of chytridiomycosis in introduced American bullfrogs terrestrial species of anurans have been observed and native amphibians in the Yunnan Province, China. with Bd, suggesting frog to frog transmission is EcoHealth. 7(1):127-134. doi: 10.1007/s10393-010-0307-0 possible (Kriger et al. 2007). Başkale E, Yildirim E,Çevik IE, Kaya U. 2013. Population Differences in morbidity and mortality in size and age structure of metamorphic and experimentally infected amphibians indicate that Bd pedomorphic forms of Ommatotriton ophryticus virulence can vary between strains of the same and (Berthold, 1846) in the Northwestern Black Sea different lineages. Increased Bd growth rate, Region of Turkey. J Herpetol. 47(2):270-276. zoospore production, and sporangial size in pure doi: 10.1670/11-116 culture, have been linked with increased host Boyle DG, Boyle DB, Olsen V, Morgan JAT, Hyatt AD. mortality and immunosuppressive activity (Fisher et 2004. Rapid quantitative detection of al. 2009). Our studies suggested that amphibians can chytridiomycosis Batrachochytrium dendrobatidis in evolve resistance to Bd and may have the ability to amphibian samples using real-time taqman PCR assay. Dis Aquat Organ. 60(2):141-148. coexist with the disease. The Eastern Black Sea doi: 10.3354/dao060141 region has unique reptile fauna. These regions are the Bülbül U, Matsui M, Kutrup B, Eto K. 2011. Taxonomic corridors of the species coming from the Caucasus relationships among Turkish water frogs as revealed and the south. The high Anatolian diagonal by phylo-genetic analyses using mtDNA gene mountains are a barrier to colonization (Ansell et al. sequences. Zool Sci. 28(12):930-936. 2011). This study showed that Anatolian diagonal doi: 10.2108/zsj.28.930 mountains are not a barrier for the colonization of Bd Carver S, Bell BD, Waldman B. 2010. Does in Anatolia. If amphibians can evolve resistance to chytridiomycosis disrupt amphibian skin function? Bd and may have the ability to coexist with the Copeia. 2010(3):487-495. disease, testing for the presence of Bd should be doi: 10.1643/CH-09-128 Daszak P, Berger L, Cunningham AA, Hyatt AD, Green mandatory other regions of Turkey. DE, Speare R. 1999. Emerging infectious diseases and In conclusion, the uncertain distribution and amphibian population declines. Emerg Infect Dis. potential impact of Bd presence in Turkey require 5(6):735-748. additional investigation before accurate evaluations doi : 10.3201/eid0506.990601 can be made. Standardized field surveillance Di Rosa I, Simoncelli F, Fagotti A, Pascolini R. 2007. methods and laboratory diagnostic techniques are Ecology: the proximate cause of frog declines? needed to more carefully investigate. The presence, Nature. 447(7144):4-5. distribution, virulence to native species and clade doi: 10.1038/nature05941 membership of Bd in Turkey must be verified before Erismis UC, Konuk M, Yoldas T, Agyar P, Yumuk D, its potential impact on Anatolian amphibians can be Korcan SE. 2014. Survey of Turkey’s endemic frogs for amphibian chytrid fungus Batrachochytrium accurately predicted. dendrobatidis. Dis Aquat Organ. 111(2):153-157. doi: 10.3354/dao02742 Acknowledgements Farrer RA, Weinert LA, Bielby J, Garner TW, Balloux F, This research was supported by Project no. Clare F, Bosch J, Cunningham AA, Weldon C, du Tubitak 113Z139. Ethical endorsement was ratified Preez LH, Anderson L, Pond SL, Shahar-Golan R, by the Ethical Committee of Afyon Kocatepe Henk DA, Fisher MC. 2011. Multiple emergences of University and the Turkish Department of Nature genetically diverse amphibian-infecting chytrids Conservation (Permit number, DKMP-51039719). include a globalized hypervirulent recombinant lineage. P Natl Acad Scı Usa. 108(46):18732-18736. References doi: 10.1073/pnas.1111915108 Anna ES, Grismer LL, Anuar S, Onn CK, Grismer JL, Fisher MC, Garner TW, Walker SF. 2009. Global Quah E, Muin MA, Ahmad N, Lenker M, Zamudio emergence of Batrachochytrium dendrobatidis and KR. 2011. First record of Batrachochytrium amphibian chytridiomycosis in space, time, and host. dendrobatidis infecting four frog families from Annu Rev Microbiol. 63:291-310. Peninsular Malaysia. Ecohealth. 8(1):121-128. doi: 10.1146/annurev.micro.091208.073435 doi: 10.1007/s10393-011-0685-y Franzen M, Bussmann M, Kordges T, Thiesmeier B. Ansell SW, Stenøien HK, Grundmann M, Russell SJ, 2008. Die Amphibien und Reptilien der Südwest- Koch MA, Schneider H, Vogel JC. 2011. The Türkei. Bielefeld: Laurenti Verlag 329 p. importance of Anatolian mountains as the cradle of Göçmen B, Veith M, Iğci N, Akman B, Godmann O, global diversity in Arabis alpina, a key arctic–alpine Wagner N. 2013. No detection of the amphibian species. Annals of Botany. 108(2):241-252. pathogen Batrachochytrium dendrobatidis in doi: 10.1093/aob/mcr134 terrestrial Turkish salamanders (Lyciasalamandra)
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despite its occurrence in syntopic frogs (Pelophylax doi: 10.1046/j.1365-294X.2003.01732.x bedriagae). Salamandra. 49(1):51-55. Richmond JQ, Savage AE, Zamudio KR, Rosenblum EB. Hyatt AD, Boyle DG, Olsen V, Boyle DB, Berger L, 2009. Toward immunogenic studies of amphibian Obendorf D. 2007. Diagnostic assays and sampling chytridiomycosis: linking innate and acquired protocols for the detection of Batrachochytrium immunity. Bioscience 59(4):311-32. dendrobatidis. Dis Aquat Organ. 73(3):175-192. doi: 10.1525/bio.2009.59.4.9 doi: 10.3354/dao073175 Ron SR. 2005. Predicting the distribution of the amphibian Kriger KM, Hines HB, Hyatt AD, Boyle DG, Hero JM. pathogen Batrachochytrium dendrobatidis in the new 2006. Techniques for detecting chytridiomycosis world. Biotropica. 37(2):209-221. in wild frogs: comparing histology with doi: 10.1111/j.1744-7429.2005.00028.x real-time taqman PCR. Dis Aquat Organ. 71(2):141- Sensoy S, Demircan M, Ulupınar Y, Balta İ. 2008. Türkiye 148. İklimi; [cited 18 Feb 2018]. Available from doi: 10.3354/dao071141 https://www.mgm.gov.tr/FILES/genel/makale/13_tur Kriger KM, Pereoglou F, Hero JM. 2007. Latitudinal kiye_iklimi.pdf variation in the prevalence and intensity of chytrid Şekercioğlu CH, Anderson S, Akçay E, Bilgin R, Emre Batrachochytrium dendrobatidis infection in eastern Can Ö, Semiz G, Tavşanoğlu C, Baki Yokeş M, Australia. Conserv Biol. 21(5):1280-1290. Soyumert A, İpekdal K, Sağlam İK, Yücel M, Dalfes doi: 10.1111/j.1523-1739.2007.00777.x HN. 2011. Turkey’s globally important biodiversity in Marantelli G, Berger L, Speare R, Keegan L 2004. crisis. Biol Conserv. 144(12):2752-2769. Distribution of the amphibian chytrid doi:10.1016/j.biocon.2011.06.025 Batrachochytrium dendrobatidis and keratin during Veith M, Schmidtler F, Kosuch J, Baran I, Seitz A. 2003. tadpole development. Pacific Conservation Biology. Paleoclimatic changes explain Anatolian mountain 10(2):173-179. frog evolution: a test for alternating vicariance and doi: 10.1071/PC040173 dispersal event. Mol Ecol. 12(1):185-189. Mazzoni R, Cunningham AA, Daszak P, Apolo A, doi: 10.1046/j.1365-294X.2003.01714.x Perdomo E, Speranza G. 2003. Emerging pathogen of Weldon C, du Preez LH, Hyatt AD, Muller R, Speare R. wild amphibians in frogs.Rana catesbeiana farmed for 2004. Origin of the amphibian chytrid fungus. Emerg international trade. Emerg Infect Dis. 9(8):995-998. Infect Dis. 10(12):2100-2105. doi: 10.3201/eid0908.030030 doi: 10.3201/eid1012.030804 Morehouse EA, James TY, GanleyA RD, Vilgalys R, Yildiz MZ, Göcmen B. 2012. Population dynamics, Berger L, Murphy PJ, Longcore J E. 2003. Multilocus reproduction, and life history traits of Taurus Frog, sequence typing suggests that the chytrid pathogen of Rana holtzi Werner, 1898 (Anura: Ranidae) in amphibians is a recently emerged clone. Mol Ecol. Karagöl (Ulukışla, Niğde), Turkey. Herpetologica 12(2):395-403. Romanica.6:1-40.
LIMNOFISH-Journal of Limnology and Freshwater Fisheries Research 5(1): 34-40 (2019)
Dietary Protein Requirements of Zebrafish (Dania rerio)
Hüseyin SEVGİLİ * , Soner SEZEN , Mahir KANYILMAZ , Özgür AKTAŞ , Faruk PAK
Mediterranean Fisheries Research Production and Training Institute, Kepez Unit, 07192, Döşemealtı, Antalya, Turkey
ABSTRACT ARTICLE INFO
Zebrafish (Danio rerio) with an initial weight of 88.61±0.82 mg were fed eight RESEARCH ARTICLE isoenergetic diets containing dietary protein levels ranging from 20 to 55 % by 5 % increments. Each diet was feed in triplicate of fish for 6 weeks. Specific growth Received : 04.07.2018 rates (SGR) at week 2 and 4 were quadratically affected by the treatments but this Revised : 22.10.2018 trend disappeared at the end of the experiment. Dietary protein levels linearly Accepted : 30.10.2018 reduced the values of daily feed intake, feed conversion ratio and protein efficiency rate. The whole body dry matter, ash and lipid concentrations linearly Published : 25.04.2019 decreased with dietary protein levels whereas whole body protein was quadratically affected. The second order polynomial and two break point linear DOI:10.17216/LimnoFish.440537 models (TBPLM) were used to estimate dietary protein requirements. The later model generated lower residual sum of squares when SGRWeek4 and SGRFinal * CORRESPONDING AUTHOR values were used as a response. Minimum dietary protein requirements for [email protected] SGRWeek4 and SGRFinal were estimated by the TBPLM as 27.69 and 28.93 % Phone : +90 242 251 0585 respectively. Briefly, results of the study suggest a minimum dietary protein requirement of zebrafish is about 29 % for maximum growth rate. Keywords: Zebrafish, dietary protein, growth, feeding
Zebra Balığının (Dania rerio) Diyetsel Protein Gereksinimi Öz: Ortalama başlangıç ağırlığı 88,61±0,82 mg olan zebra balıkları (Danio rerio) protein düzeyi % 20-55 arasında değişen sekiz adet izoenerjitik yemle beslenmiştir. Her bir deneme yemi üç tekrarlı olarak 6 hafta boyunca balıklara verilmiştir. Spesifik büyüme oranı (SGR) 2. ve 4. haftalarda kuadratik olarak etkilenirken, bu eğilim deneme sonunda kaybolmuştur. Protein düzeyleri arttıkça yem tüketimi, yemden yararlanma oranı ve protein etkinlik oranı doğrusal olarak düşmüştür. Tüm vücut kuru madde, kül ve lipit konsantrasyonları diyetsel protein düzeyinin artışı ile doğrusal olarak düşmüş, vücut protein düzeyi ise kuadratik olarak etkilenmiştir. Diyetsel protein gereksinimlerini tahmin etmek için, ikinci derece regresyon ve iki kırıklı linear model (İKLM) kullanılmıştır. 4. hafta ve deneme sonu SGR değerleri kullanıldığında İKLM daha düşük kalıntı kareler toplamı vermiştir. 4. hafta ve final SGR oranlarına göre, İKLM minimum protein gereksinimlerini sırasıyla, % 27,69 ve % 28,93 olarak tahmin etmiştir. Kısaca, çalışma bulguları zebra balıklarının maksimum büyüme için minimum protein gereksinimlerinin yaklaşık % 29 olduğunu göstermektedir. Anahtar kelimeler: Zebra balığı, diyetsel protein, büyüme, yemleme
How to Cite Sevgili H, Sezen S, Kanyılmaz M, Aktaş Ö, Pak F. 2019. Dietary Protein Requirements of Zebrafish (Dania rerio). LimnoFish. 5(1): 34-40. doi: 10.17216/LimnoFish.440537
Introduction al. 2007; Siccardi III et al. 2009; Smith Jr et al. 2013), Zebrafish is used in a wide range of scientific biotin requirements (Yossa et al. 2014) and effects of disciplines as a model animal. Basic culture dietary carbohydrate levels on growth and nutrient requirements particularly nutritional needs of utilization performance and hepatic transcriptome by zebrafish however are still incomplete (Lawrence sexes (Robison et al. 2008), although there are some 2007; Ulloa et al. 2014). Existing literature about others. zebrafish nutrition has dealt with some topics Dietary protein level in fish is considered as one including the evaluation several diet types and of most important criterions since it is most the protein sources (Artemia, paste liver, flake, expensive nutrient and affects a number of functions commercial trout and experimental diets) in terms of from molecular level to growth related traits reproductive and growth performance (Markovich et (Lawrence 2007; NRC 2011; Ulloa et al. 2011; Ulloa
Sevgili et al. 2019 - LimnoFish 5(1): 34-40 35 et al. 2014). Despite its fundamental importance in sacrificed by an overdose of ethylene glycol nutritional physiology, dietary protein requirement of monophenyl ether (1.2 mL/L) for final proximate juvenile zebrafish has been studied recently by analysis. Fernandes et al. (2016), who estimated the minimum Experimental diets dietary requirements between 37.6 and 44.8% for Diets were formulated based on dry matter basis maximum weight gain and protein retention using a using the linear method in Winfeed 2.8 (Winfeed four-parameter saturation kinetics model (SKM) and Ltd., Cambridge, UK). Eight isoenergetic diets (18 broken line model (BLM). O'Brine et al. (2015) also MJ/kg gross energy (GE)) were formulated to studied protein and lipid requirements of older provide crude protein (CP) levels from 20 to 55 % by zebrafish (ca. 4 months) and reported using ANOVA 5 % increments (Table 1). The dietary protein level that diet with 32% dietary and 8% lipid can be was increased by adjusting the fraction of the fish sufficient for growth. Growth rate of zebrafish can meal in the diet. Fish meal was used were used as vary greatly by laboratories, populations and batches primary protein source whereas a 1:1 mixture of (Eaton and Farley 1974), plus the estimations dietary soybean meal and corn gluten meal was used as requirements of fish are subjected to huge variations secondary protein source. Wheat starch and due to the selected statistical model and response sunflower oil served as carbohydrate and lipid variables (Hernandez-Llamas 2009; NRC 2011). sources, respectively. Therefore, a six-week feeding trial with juvenile All the dietary ingredients were ground with a zebrafish from 42 to 84 days post hatching was hammer mill (Kocamaz Machine, Model KT-20C, planned to estimate dietary protein requirements. İzmir, Turkey), weighed at predetermined levels, thoroughly mixed and then extruded into 2 mm using Materials and Methods a pasta machine (model P3, La Monferrina, Italy). Fish and rearing system The resulting material was air dried at a room. The experiment was carried out at the Kepez Unit of Mediterranean Fisheries Research Production and Calculation and chemical analysis Training Institute, Antalya, Turkey. A total of 720, Daily feed intake (DFI g/kg MBW / day) = (dry matter 35day post hatching zebrafish (pink type) were intake / MBW0.8) / day randomly allocated in groups of 30 across 24, 10L Metabolic body weight (MBW) = (Geometric mean tanks. Fish were acclimated for a week and fed a of initial weight (IW) and final weight (FW))0.8 commercial rainbow trout diet with 60 % protein and 10 % lipid and 150-300 µm particle diameter Specific growth rate (SGR) = 100 × [(ln FW – ln (Bioaqua, Çamlı Yem, İzmir, Turkey). The average IW)/day individual weight per tank was 88.61 ± 0.82 mg and Daily feed intake (mg/kg MBW0.8/day-1) = (dry feed the age was 6 weeks. intake / MBW0.8) / days The experimental tanks were connected to a recirculation system. Daily water renewal rate of the Feed conversion ratio (FCR) = dry matter intake / system was 30 %. Each tank was given 100 mL/min weight gain of water and provided with aeration using one air Protein efficiency ratio (PER) = weight gain / protein stone. Average water temperature, oxygen, pH, NH4- fed N and NO2-N concentrations in the system over the 0.8 -1 experiment were checked twice a week and were Daily nutrient intake (g/kg MBW /day ) = [(protein, 0.8 24.87±0.49°C, 7.65±0.06 mg/L, 8.52±0.06, <0.02 energy intake / MBW ) / days. mg/L and 0.013±0.003 mg/L, respectively. A natural Daily nutrient gain (g/kg MBW0.8/day) = [(final body photoperiod was applied as 13-14 h L: 11-10 h D. weight ×final body nutrient) – (initial body weight × Fish were biweekly weighed in bulk after an initial body nutrient)] / MBW0.8 / days. anesthetization with ethylene glycol monophenyl ether (0.3 mL/L). Feed was withheld on the weighing Nutrient retention (%) = 100 × (daily nutrient gain / days. Feed particle diameters were 300-500, 500-800 daily nutrient intake). and 800-1000 µm during 0-2, 2-4 and 4-6 weeks of Fish samples were stored at -20 °C until analysis. the experiment. Fish were fed ad libitum by hand Prior to analysis, they were chopped into very tiny twice a day at 09:00 and 16:00 h. Each feed was tried pieces using knife. Proximate analysis, except crude in triplicated tanks and was carefully administered lipid, of experimental diets and fish were performed until the feeding activity ceased. At the start of the according to the methods of AOAC (1990): dry experiment, a composite sample of five fish per tank matter at 104 °C till constant weight, ash content by were taken for initial body composition whereas at incineration in a muffle furnace at 600 °C for 2 h; CP the end of the experiment, all fish per tank were (N×6.25) by the Kjeldhal method after acid digestion.
36 Sevgili et al. 2019 - LimnoFish 5(1): 34-40
Lipid was determined with ether-extraction using an the model fitting best in terms of the residual sum of automatic extraction system (ANKOMXT15 squares (Hernandez-Llamas 2009). Extractor, ANKOM Technology, Macedon, USA). The equations of second order polynomial Statistical analysis regression (1) and TBPLM (2, 3 and 4) are given Polynomial contrasts were used to detect linear below. and quadratic effects of dietary protein levels on the 2 푦 = 푖1 + 푏1푥 + 푏2푥 (1) observed response variables. Significant treatment effects were considered at P≤0.10. Statistical where 푖1is intercept, b1 and b2 is are the analyses were conducted in JMP v.8.0 (SAS Institute regression coefficients (Shearer 2000). Inc. 2008). To estimate dietary protein requirements 푦 = 푖1 + 푏1푥 if 푥 < 푥bp, (2) for average SGRWeek4 and SGRFinal, two models were tested using GRAPHPAD PRISM 5 for Windows 푦 = 푦푚푎푥 + 푏2푥 if 푥 ≥ 푥bp, (3) (GraphPad Software, San Diego, CA, USA): second 푦 = 푖 + 푏 푥 if 푥 > 푥 (4) order polynomial regression and two-break points 2 3 bp non-linear model (TBPLM). The latter is a where i1 and b1 are parameters describing the combination of conventional broken line model positive linear relation, ymax is the maximum response (Hernandez-Llamas 2009) with a negative linear and i2 and b3 are parameters of negative linear regression at the right side of the response curve. The relation. To assume a constant response, the slope at optimum dietary protein levels were defined based on the plateau (b2) was set at zero. Table 1. Formulation and nutrient composition of experimental diets (% dry matter) Ingredients 20P 25P 30P 35P 40P 45P 50P 55P Fish meal 19.87 26.49 30.90 37.51 41.92 48.54 52.95 59.56 Soybean meal 2.14 3.06 3.67 4.59 5.20 6.12 6.73 7.65 Corn gluten meal 2.14 3.06 3.67 4.59 5.20 6.12 6.73 7.65 Wheat starch (Cooked) 64.15 54.83 48.61 39.28 33.07 23.74 17.53 8.20 Sunflower oil 8.21 7.53 7.08 6.40 5.94 5.26 4.81 4.13 MCP1 2.24 2.07 1.96 1.80 1.68 1.52 1.41 1.24 Mineral mixture2 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Vitamin mixture3 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Choline chloride 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 CMC4 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Alpha cellulose5 0.00 1.72 2.87 4.59 5.73 7.45 8.60 10.32 Nutrient levels (% dry matter) Dry matter 91.41 91.74 91.64 91.89 92.31 92.24 92.66 92.49 Crude ash 5.75 6.73 6.94 7.63 8.21 8.98 9.71 10.51 Crude lipid 10.26 10.02 10.32 10.94 10.62 9.97 10.28 10.34 Crude protein 20.38 26.22 28.93 34.94 39.56 44.20 49.72 56.88 Gross energy (MJ/kg) 19.82 19.69 19.70 19.81 19.74 19.47 19.58 19.62 Protein energy ratio (g/MJ) 10.28 13.32 14.69 17.64 20.04 22.70 25.39 28.99
Results (linear, P=0.666 and quadratic, P=0.213) (Table 2). All experimental groups more than tripled Dietary protein levels had a strong linear effect on their initial weights during the 6-week daily feed intake, FCR and PER (linear, P=<0.001). experiment (Table 2). There was a weak The second order polynomial model generated quadratic effect of dietary protein levels on 4th week 0.0148 and 0.0293 of residual sum of squares for weight (quadratic, P=0.104) but it disappeared SGRWeek4 and SGRFinal respectively, whereas the at the final. SGR values at week 2 and 4 TBPLM yielded lower levels with 0.0128 and 0.0248. were quadratically affected by the treatments Minimum dietary protein requirements for SGRWeek4 (quadratic, P=0.025 and P=0.060 respectively), and SGRFinal were estimated by the TBPLM as 27.69 which also vanished at the end of the experiment and 28.93% respectively (Figure 1).
Sevgili et al. 2019 - LimnoFish 5(1): 34-40 37
Table 2. Growth, and nutrient utilization performance of zebrafish fed varying dietary protein levels W at 4th W at 2nd SGR at SGR at 4th SGR at Daily feed IW week W at final Diets week 2nd week week final intake (g/ kg- FCR (mg/ fish) (mg/ (mg/fish) PER (mg/fish) (%/day) (%/day) (%/day) MBW0.8 /day) fish) 20P 86.81 117.70 195.29 293.86 2.34 2.90 2.90 42.74 2.00 2.46
25P 88.32 122.92 203.80 300.03 2.54 2.99 2.91 38.70 1.81 2.12
30P 86.36 123.64 203.25 310.87 2.76 3.06 3.05 37.66 1.66 2.08
35P 88.61 122.80 205.94 312.20 2.50 3.01 3.00 34.89 1.56 1.83
40P 90.25 126.40 208.53 320.32 2.60 3.00 3.02 32.53 1.44 1.76
45P 88.64 120.27 197.80 291.18 2.35 2.87 2.83 28.44 1.37 1.66
50P 88.36 120.66 201.11 306.55 2.39 2.94 2.96 29.55 1.35 1.50
55P 91.57 120.73 198.78 306.74 2.13 2.77 2.88 24.80 1.16 1.52
Pooled 2.532 3.512 5.647 10.08 0.152 0.094 0.075 0.764 0.053 0.059 SEM
Linear 0.768 0.844 0.471 0.109 0.170 0.666 <0.001 <0.001 <0.001 P values Quadratic 0.136 0.104 0.277 0.025 0.060 0.213 0.114 0.017 <0.001 IW; initial weight, W; weight, SGR; specific growth rate, MBW, metabolic body weight, FCR; feed conversion rate, PER; protein efficiency ratio, Pooled SEM, standard error of the means.
Figure 1. Effect of dietary protein levels on SGRWeek4 and SGRFinal values in zebrafish. Values are represented as the mean SEM of three replicates. SGR; specific growth rate. The whole body dry matter, ash and lipid (linear, P=<0.001; quadratic, P=<0.010) (Table 4). concentrations linearly decreased with dietary On the other hand, no effect of dietary protein levels protein levels (P= <0.001) whereas whole body was observed on daily protein gain. Dietary protein protein was quadratically affected (P=0.050) levels linearly decreased daily energy gain and (Table 3). energy retention of zebrafish (linear, P=<0.001), Daily protein and energy intakes by zebrafish whereas quadratically decreased protein retention quadratically decreased in response to dietary protein (linear and quadratic, P=<0.001).
38 Sevgili et al. 2019 - LimnoFish 5(1): 34-40
Table 3. Whole body compositions of zebrafish fed varying levels of dietary protein (%) Diets Whole body dry matter Whole body ash Whole body lipid Whole body protein Initial 27.01 2.79 7.49 14.49 20P 30.95 3.09 9.82 16.14 25P 30.43 3.03 9.08 16.38 30P 29.96 3.09 8.55 16.14 35P 29.69 2.73 8.31 15.77 40P 30.09 2.92 8.24 16.37 45P 28.77 2.69 7.09 15.77 50P 28.55 2.62 7.07 16.17 55P 28.95 2.78 7.11 16.86
Pooled SEM 0.351 0.084 0.296 0.302
Linear <0.001 <0.001 <0.001 0.433 P values Quadratic 0.227 0.117 0.049 0.05
Pooled SEM; standard error of the means Table 4. Nutrient utilization of zebrafish fed graded levels of dietary protein Protein Energy Daily protein intake Daily energy intake Daily protein gain Daily energy gain Diets retention retention (g/kg MBW0.8/day) (kJ/kg MBW0.8/day) (g/kg MBW0.8/day) (kJ/kg MBW0.8/day) (%) (%) 20P 8.71 926.89 3.60 176.29 41.42 19.06
25P 10.15 830.39 3.70 170.57 36.45 20.54
30P 10.89 809.35 3.81 170.57 34.96 21.09
35P 12.19 752.09 3.64 162.44 29.84 21.61
40P 12.87 695.66 3.87 167.92 30.10 24.17
45P 12.57 600.37 3.40 137.56 27.04 22.92
50P 14.69 624.19 3.71 148.02 25.34 23.79
55P 14.11 526.05 3.82 149.34 27.11 28.41
Pooled SEM 0.272 16.379 0.156 7.044 1.578 1.153
P Linear <0.001 <0.001 0.744 <0.001 <0.001 <0.001 values Quadratic 0.001 0.074 0.76 0.471 <0.001 0.317 Pooled SEM; standard error of the means, MBW; metabolic body weight Discussion the study with a significant quadratic trend, but with The responses of zebrafish to dietary protein a lower rate during the later periods. This could be levels in the present study displayed some differences resulted from that the fish were not able to totally from those of the previous studies (Fernandes et al. adapted to the experimental conditions even after a 2016; O'Brine et al. 2015). This could be resulted week of acclimation period. The SGR responses were from several factors including growth depensation in abated but with still a significant quadratic trend at 4th zebrafish, differences in strain and in number of sexes week, and became insignificant at the final, in experimental tanks and maturational stages as suggesting a decrease at the intensity of growth underlined previous authors (Biga and Goetz 2006; response with ages to dietary protein level. Although Eaton and Farley 1974). Since we did not define difficult to compare the results of this study with maturational situation and sexes of the individuals in those of O'Brine et al. (2015) who used a higher range the present study, we were unable to conclude their of dietary protein levels between 32 and 75%, no contributions to the differences in our results and significant treatment effect on growth rate of about 4- those of O'Brine et al. (2015) and Fernandes et al. month-old zebrafish was determined. The impacts of (2016). developmental stages on zebrafish growth rate has SGRs of zebrafish reared on increasing levels of been previously underlined (Eaton and Farley 1974). nd dietary protein were affected as early as 2 week of Yet, we used SGRWeek4 and SGRFinal values as
Sevgili et al. 2019 - LimnoFish 5(1): 34-40 39 response variables to estimate the dietary protein levels in zebrafish laboratories. Our results related requirements. The TBPLM estimated the with FCR showed a quadratic decrease in response to requirements for SGRWeek4 and SGRFinal as 27.69 and the increase in dietary protein, being consistent to a 28.93% respectively without a considerable change certain degree with those of Fernandes et al. (2016), with fish size. Dietary protein requirement levels of who observed an improvement in feed efficiency up zebrafish estimated here are consistent with those of to 35% protein level, then a plateau. omnivorous species such as common carp and A quadratic decrease in PER with increasing goldfish reported by NRC (2011) and Ulloa et al. dietary protein level was the case in the present study. (2011). But, our findings are lower than those levels This suggests that zebrafish did not use increasing of 37.6 and 44.8 % for zebrafish by Fernandes et al. dietary protein particularly at above requirement (2016), who used average estimated values of SKM levels for protein synthesis as indicated several fish and BLM based on weight gain and protein retention. species including Arctic charr, Salvelinus alpinus The model with two breaks used in the present study (Gurure et al. 1995), Zacco barbata (Shyong et al. was previously employed by Klatt et al. (2016) for 1998), marbled spinefoot rabbitfish, Siganus estimation of lower and upper critical dietary rivulatus (El‐ Dakar et al. 2011) and tiger puffer, concentrations of methionine+cysteine for juvenile Takifugu rubripes (Kim and Lee 2009). turbot (Psetta maxima). The second order polynomial The effect of dietary protein levels on whole body model is widely used in estimation of nutrient compositions of zebrafish was a significant linear requirements of aquaculture species (Shearer 2000), decrease in dry matter, crude ash and lipid whereas but the TBPLM fitted better in the present study in no change in crude protein in the present study. Our terms of residual sum of squares, suggesting that it dry matter results are consistent with those of can be used in future studies as an alternative model Fernandes et al. (2016), but this was not the case in for determination of minimum nutrient requirements. the whole body protein which displayed an increase When it comes to right side of the curve, the present with dietary protein levels in their study. Although no model estimated an inhibition dietary protein level of clear consensus about the effects of dietary protein 39.56%. However, since the right side of the curve levels on the proximate compositions of fish in the did not display a clear descending trend, a great literature, Gurure et al. (1995) found a decrease in dry caution should be exercised before a definite matter and crude lipid concentrations in Arctic charr conclusion is reached in terms of inhibition level of with dietary protein levels, being fully in parallel dietary protein. The descending trend at the right side with our findings. Higher lipid concentrations in of SGRs curve is inconsistent with previous zebrafish on lower dietary protein levels could be a observations in zebrafish (Fernandes et al. 2016; result of higher depositions of energy due to higher O'Brine et al. 2015), who found a plateau at high feed consumption. protein levels. We can conclude that our SGR data Expectedly, daily protein intake of zebrafish appears to be suitable for estimation of only increased with dietary protein level. Similar results minimum dietary protein using the TBPLM model were also recorded by other authors in different fish level but not for the inhibition level. Yet, care should species including zebrafish (Akpιnar et al. 2012; El‐ be exercised that dietary protein levels above about Dakar et al. 2011; Fernandes et al. 2016). However, 45% may lead to a reduction in growth performance this trend was not reflected to daily protein gain, of juvenile zebrafish, at least in the studied weight which in turn resulted in a significant quadratic ranges. decrease in protein retention in response to increasing Feed consumption of fish linearly decreased with levels of protein as was the case in PER values. the increase of dietary protein levels. This is Although the protein retention data are in harmony consistent with the results reported by Akpιnar et al. with those by Fernandes et al. (2016) at a certain (2012) and Fernandes et al. (2016), who observed an degree, daily protein gains are inconsistent with the inverse relation between feed intake and dietary findings of these authors. Our daily energy intake and protein in juvenile shi drum (Umbrina cirrosa) and gain values displayed a linear decrease with dietary zebrafish. This phenomenon could be attributed to protein levels but energy retention showed an inverse compensatory response to get more protein in fish fed trend, being partly in parallel with the results of lower dietary protein levels, as argued by several Fernandes et al. (2016). authors (Akpιnar et al. 2012; El‐ Dakar et al. 2011; In conclusion, the results of the present Fernandes et al. 2016; O'Brine et al. 2015). experiment show that zebrafish growing from 85 and Therefore, at restricted feeding regimes at the 300 mg require minimum 29% dietary protein estimated requirement level in this study fish level in their diets including about 10% lipid or 19.5 may not meet their daily protein requirements MJ/kg gross energy when fed ad libitum. Further and significant attention should be paid to feeding studies are required to determine the effects of
40 Sevgili et al. 2019 - LimnoFish 5(1): 34-40 varying dietary protein to energy ratios at different rerio): a review. Aquaculture. 269(1-4):1-20. feeding levels. doi: 10.1016/j.aquaculture.2007.04.077 Markovich ML, Rizzuto NV, Brown PB. 2007. Diet Acknowledgements affects spawning in zebrafish. Zebrafish. 4(1):69-74. doi: 10.1089/zeb.2006.9993 The authors would like to thank Fikri Çağlar NRC. 2011. Nutrient requirements of fish and shrimp. Yücel, Hamza Toprak and Ramazan Dolaşık for their National Research Council of the National help during the experiment. Academies, Washington DC. O'Brine TM, Vrtělová J, Snellgrove DL, Davies SJ, References Sloman KA. 2015. Growth, oxygen consumption, and Akpιnar Z, Sevgili H, Özgen T, Demir A, Emre Y. 2012. behavioral responses of Danio rerio to variation in Dietary protein requirement of juvenile shi drum, dietary protein and lipid levels. Zebrafish 12(4):296- Umbrina cirrosa (L.). Aquacult Res. 43:421-429. 304. doi: 10.1111/j.1365-2109.2011.02845.x doi: 10.1089/zeb.2014.1008 AOAC. 1990. Official Methods of Analysis, 15 ed., Robison BD, Drewa RE, Murdoch GK, Powell M, Association of Official Analytical Chemists, Rodnick KJ, Settles M, Stone D, Churchill A, Hill RA, Arlington, VA, p.684 Papasani MR, Lewis SS, Hardy RW. 2008. Sexual Biga PR, Goetz FW. 2006. Zebrafish and giant danio as dimorphism in hepatic gene expression and the models for muscle growth: determinate vs. response to dietary carbohydrate manipulation in the indeterminate growth as determined by morphometric zebrafish (Danio rerio). Comp Biochem Physiol Part analysis. Am J Physiol Regul Integr Comp Physiol. D Genomics Proteomics. 3(2):141-154. 291:R1327-R1337. doi: 10.1016/j.cbd.2008.01.001 doi: 10.1152/ajpregu.00905.2005 Shearer K. 2000. Experimental design, statistical analysis Eaton RC, Farley RD. 1974. Growth and the reduction of and modelling of dietary nutrient requirement studies depensation of zebrafish, Brachydanio rerio, reared in for fish: a critical review. Aquacult Nutr. 6(2):91-102. the laboratory. Copeia. 1974(1):204-209. doi: 10.1046/j.1365-2095.2000.00134.x doi: 10.2307/1443024 Shyong W-J, Huang C-H, Chen H-C. 1998. Effects of El‐ Dakar AY, Shalaby SM, Saoud IP. 2011. Dietary dietary protein concentration on growth and muscle protein requirement of juvenile marbled spinefoot composition of juvenile Zacco barbata. Aquaculture. rabbitfish Siganus rivulatus. Aquacult Res. 42:1050- 167:35-42. 1055. doi: 10.1016/S0044-8486(98)00313-5 doi: 10.1111/j.1365-2109.2010.02694.x Siccardi III AJ, Garris HW, Jones WT, Moseley DB, Fernandes H, Peres H, Carvalho AP. 2016. Dietary protein D'Abramo LR, Watts SA. 2009. Growth and survival requirement during juvenile growth of Zebrafish of zebrafish (Danio rerio) fed different commercial (Danio rerio). Zebrafish. 13(6):548-555. and laboratory diets. Zebrafish. 6(3):275-280. doi: 10.1089/zeb.2016.1303 doi: 10.1089/zeb.2008.0553 Gurure R, Moccia R, Atkinson J. 1995. Optimal protein Smith Jr DL, Barry RJ, Powell ML, Nagy TR, D'Abramo requirements of young Arctic charr (Salvelinus L, Watts SA. 2013. Dietary protein source influence alpinus) fed practical diets. Aquacult Nutr. 1:227-234. on body size and composition in growing zebrafish. doi: 10.1111/j.1365-2095.1995.tb00048.x Zebrafish. 10(3):439-446. Hernandez-Llamas A. 2009. Conventional and alternative doi: 10.1089/zeb.2012.0864 dose–response models to estimate nutrient Ulloa PE, Iturra P, Neira R, Araneda C. 2011. Zebrafish as requirements of aquaculture species. Aquaculture. a model organism for nutrition and growth: towards 292:207-213. comparative studies of nutritional genomics applied to doi: 10.1016/j.aquaculture.2009.04.014 aquacultured fishes. Rev Fish Biol Fisheries Kim S-S, Lee K-J. 2009. Dietary protein requirement of 21(4):649-666. juvenile tiger puffer (Takifugu rubripes). Aquaculture. doi: 10.1007/s11160-011-9203-0 287(1-2):219-222. Ulloa PE, Medrano JF, Feijoo CG. 2014. Zebrafish as doi: 10.1016/j.aquaculture.2008.10.021 animal model for aquaculture nutrition research. Front Klatt SF, von Danwitz A, Hasler M, Susenbeth A. 2016. Genet. 5:313. Determination of the lower and upper critical doi: 10.3389/fgene.2014.00313 concentration of Methionine+ Cystine in diets of Yossa R, Sarker PK, Mock DM, Vandenberg GW. 2014. juvenile turbot (Psetta maxima). Aquaculture. 452:12- Dietary biotin requirement for growth of juvenile 23. zebrafish Danio rerio (Hamilton-Buchanan). doi: 10.1016/j.aquaculture.2015.10.015 Aquacult Res. 45(11):1787-1797. Lawrence C. 2007. The husbandry of zebrafish (Danio doi: 10.1111/are.12124
LIMNOFISH-Journal of Limnology and Freshwater Fisheries Research 5(1): 41-46 (2019)
Zooplankton Fauna of Abant Lake: Past and Present
Pınar GÜRBÜZER 1* , Ezgi TÜZÜN TERESHENKO 2 , Ahmet ALTINDAĞ3 , Seyhan AKISKA3
1 Sinop University, Faculty of Fisheries, TR57000-Sinop, Turkey 2 Ankara University, Graduate School of Naturel and Applied Sciences, TR06110-Ankara, Turkey 3Ankara University, Faculty of Science, TR06100-Ankara, Turkey
ABSTRACT ARTICLE INFO
Observing the changes of zooplanktonic organisms over the years can give us RESEARCH ARTICLE consistent information about the limnological conditions of the present and future. Our aim in this study was to determine current conditions of zooplankton fauna Received : 27.07.2018 of Abant Lake, which was studied seasonally, and could provide resources for Revised : 07.11.2018 future studies. The samples of zooplankton were collected horizontally and Accepted : 08.11.2018 vertically from Abant Lake seasonally, between 2015 -2016 from seven stations. At the end of the study, a total of forty-nine zooplankton species were identified. Published : 25.04.2019 Of these, 33 belonged to Rotifera, 14 to Cladocera and 2 to Copepoda. In addition, twelve species of Rotifera, and six species of Cladocera were found as new DOI:10.17216/LimnoFish.448525 records for the Abant Lake. * CORRESPONDING AUTHOR Keywords: Rotifera, Cladocera, Copepoda, Abant Lake [email protected] Phone : +90 368 287 62 54 Abant Gölü Zooplankton Faunası: Geçmiş ve Bugün Öz: Zooplanktonik organizmaların yıllar içerisindeki değişimlerini izlemek, gölün bugünkü ve gelecekteki limnolojik durumu hakkında tutarlı bilgiler verebilir. Bu çalışmada ki amacımız, Abant Gölü’nün zooplankton faunasının güncel durumunu belirlemek ve gelecekteki çalışmalara katkı sağlamaktır. Zooplankton örnekleri, Abant Gölü’nden 2015-2016 yılları arasında mevsimsel olarak, farklı habitatlarda ki yedi istasyondan yatay ve dikey olarak alınmıştır. Çalışmanın sonunda 33 tür ile Rotifera, 14 tür ile Cladocera ve 2 tür ile Copepoda grubuna ait toplam kırk dokuz zooplankton türü tespit edilmiştir. Bu çalışma ile birlikte, 12 Rotifera türü ile 6 Cladocera türü Abant Gölü zooplankton faunası için yeni kayıt olarak bulunmuştur. Anahtar kelimeler: Rotifera, Cladocera, Copepoda, Abant Gölü
How to Cite Gürbüzer P, Tüzün Tereshenko E, Altındağ A, Akıska S, 2019. Zooplankton Fauna of Abant Lake: Past and Present. LimnoFish. 5(1): 41-46. doi: 10.17216/LimnoFish.448525
Introduction It is known that zooplankton distribution, Lake studies provide a practical and richness, and composition are affected by useful structure for community studies because biotic parameters like the presence of predators, they can give important information to trophic structure, habitat differences etc. and the researchers about climate change and abiotic parameters such as temperature, ecosystem structure etc. (Olden et al. 2006). salinity, etc. (Kaya et al. 2010; Gürbüzer et al. 2017). Zooplanktonic organisms include both predator and For example, one of the most reliable predictors prey organisms. They are primary consumers in of eutrophic condition is total phosphorus aquatic ecosystems, especially in lakes and constitute (Filstrup and Downing 2017). It is known to affect a major food source for their predator like species richness in zooplankton communities macroinvertebrates, fish and birds. Zooplankton where richness decreases with an increasing considered as indicators in lakes because of their total phosphate (Jeppesen et al. 2000). Since pivotal role in aquatic food webs (Jeppesen et al. rotifers are opportunistic species they become 2011). dominant taxa when water quality deteriorates
42 Gürbüzer et al. 2019 - LimnoFish 5(1): 41-46
(Gannon and Stremberger 1978). It is also thought Özdemir Mis et al. 2017. Our aim in this study was that observing the changes of zooplanktonic to determine current conditions of zooplankton fauna organisms over the years can give us consistent of Abant Lake, which was studied seasonally, and information about the limnological conditions of the could provide resources for future studies. present and future. Abant Lake is located in the west coast of the Materials and Methods Black Sea Region and west of the city Bolu. It has The samples of zooplankton were collected been declared as a National Park in 1988 by The from Abant Lake seasonally, between 2015 -2016 Ministry of Culture and Tourism. The area has warm from seven stations in different points (Figure 1) temperate, fully humid and warm summer according and coordinates of the sampling stations were given to Köppen-Geiger Climate Classification (Kottek et in Table 1. Samples were collected horizontally al. 2006). It is located at 1340 m above sea level, and and vertically using Hydro-Bios Plankton Net surface area of lake is 125 ha (Akşıray 1959, Erinç et (mesh size 55µ and 25 cm in diameter) and al 1961) and the maximum depth is 18 m (Çelekli and immediately fixed with 4% formalin. Külköylüoğlu 2006). Abant Lake gets attention of To identify zooplankton species, various scientists because the lake and its surroundings have resources were used which included the studies of the rich flora and fauna (Dügel et al. 2008, Karakaya Ward and Whipple (1945), Kolisko (1974), Koste et al. 2011, Atıcı and Tokatlı 2014). (1978), Harding and Smith (1974), Nogrady and There are available several zooplankton studies Pourriot (1995), Segers (1995), De Smet (1996), and about Abant Lake and the first one was conducted Smirnov (1996). In the present study, all the in 1970 by Margaritora and Cottarelli which identified taxa were checked from several checklists was later followed by extensive studies of Altındağ from Ustaoğlu 2004, Ustaoğlu et al. 2012, and (1999), Altındağ and Yiğit (2000) and Ustaoğlu 2015.
Figure 1. Study area and sampling stations Table 1. Coordinates of the sampling stations Rotifers Kellicottia longispina and Keratella Station Number Coordinates cochlearis were found during all seasons, while STA 1 40° 36' 31''N / 31° 17' 21''E Asplanchna priodonta, Synchaeta pectinata were STA 2 40° 36' 40''N / 31° 16' 55''E found during three seasons and other taxa were STA 3 40° 36' 34''N / 31° 16' 39''E identified only during two seasons or less. Cladocerans were seen during all seasons except STA 4 40° 36' 6''N / 31° 16' 22''E winter period, Alona guttata, Bosmina longirostris, STA 5 40° 35' 59''N / 31° 16' 48''E Chydorus sphaericus and Daphnia longispina were STA 6 40° 36' 15''N / 31° 17' 00''E found during three seasons. Copepods were STA 7 40° 36' 24''N / 31° 16' 55''E represented by two taxa Acanthodiaptomus denticornis and Megacyclops viridis and A. Results denticornis was found during all seasons except In this study, a total of forty-nine zooplankton winter period like cladocerans. species were identified. Of these, 33 belonged to When seasonal zooplankton changes were Rotifera, 14 to Cladocera and 2 to Copepoda. The considered it was seen that, species richness was the species list of Abant Lake was presented in Table 2. highest in autumn (twenty-five taxa), followed by 12 species of Rotifera and 6 species of Cladocera are summer and spring (twenty-four and twenty-three new records for the Abant Lake. taxa, respectively). In the winter season, only six
Gürbüzer et al. 2019 - LimnoFish 5(1): 41-46 43 species were observed all of which belonged to the analyzed, rotifers were found to have the highest taxa phylum Rotifera. percentage with 67%, while copepods had the lowest When zooplankton taxon frequencies were one with 4%.
Table 2. Species list of Abant Lake, past and present studies Species Present Study ROTIFERA Winter Spring Summer Fall MC1970 A1999 AY2000 ÖM2017 Ascomorpha ecuadis (Petry, 1850) + + Asplanchna girodi Guerne, 1888 + + + + Asplanchna priodonta Gosse, 1850 + + + + Cephalodella gibba (Ehrenberg, 1830)* + Collotheca mutabilis (Hudson, 1885)* + Collotheca ornata (Ehrenberg, 1830) + + + + Collotheca pelagica (Rousselet, 1893) + + + Collotheca sp. + Colurella adriatica Ehrenberg, 1831 + + Conochilus hippocrepis (Schrank, 1803) + + + + + Conochilus unicornis (Rousselet, 1892) + + + + Euchlanis dilatata (Ehrenberg, 1830) + + + Filinia longiseta (Ehrenberg, 1834) + + + Filinia terminalis (Plate, 1886) + + Gastropus stylifer Imhof, 1891 + + + + Kellicottia longispina (Kellicott, 1879) + + + + + + + + Keratella cochlearis (Gosse, 1851) + + + + + + + + Keratella quadrata (Müller, 1786) + + + + + + Keratella tropica (Apstein, 1907)* + Lecane bulla (Gosse, 1851) + + Lecane closteracerca (Schmarda, 1859)* + + Lecane hamata (Stokes, 1896) + + Lecane luna (Müller, 1776) + Lecane lunaris (Ehrenberg, 1832) + + + + Lecane stenroosi (Meissner, 1908)* + Lepadella acuminata (Ehrenberg, 1834)* + Lepadella ovalis (Müller, 1786)* + Lepadella patella (Müller, 1773)* + Lophocharis salpina (Ehrenberg, 1834) + + + Mytilina ventralis (Ehrenberg, 1830)* + + Notholca squamula (Müller, 1786) + + Philodina megalotrocha Ehrenberg, 1832* + Platyias quadricornis (Ehrenberg, 1832) + Polyarthra dolicoptera Idelson, 1925 + + + Polyarthra vulgaris Carlin, 1943 + + + + Squatinella mutica (Ehrenberg, 1832)* + Synchaeta litoralis Rousselet, 1902 + + Synchaeta pectinata Ehrenberg, 1832 + + + + + + Trichotria pocillum (Müller, 1776) + + + Trichotria tetractis (Ehrenberg, 1830)* +
44 Gürbüzer et al. 2019 - LimnoFish 5(1): 41-46
Table 2. Continued Species Present Study CLADOCERA Winter Spring Summer Fall MC1970 A1999 AY2000 ÖM2017 Acroperus harpae (Baird, 1843) + Alona affinis (Leydig, 1860) + Alona costata Sars, 1862 + Alona guttata Sars, 1862* + + + Alonella excisa (Fischer, 1854) + Alonella exigua (Lilljeborg, 1853) + Alonella nana (Bairs, 1850) + + Bosmina longirostris (Müller, 1785) + + + + + Ceriodaphnia quadrangula (Müller, 1785) + + + Ceriodaphnia reticulata (Jurine, 1820)* + Coronatella rectangula Sars, 1862* + Chydorus sphaericus (O.F. Müller, 1776) + + + + Daphnia hyalina Leydig 1860 + Daphnia longispina (O.F. Müller) + + + + + Diaphanosoma brachyurum (Lievin, 1848) + + + Macrothrix hirsuticornis Norman & Brady, 1867* + + Macrothrix laticornis (Jurine, 1820)* + Pleuroxus truncatus (O.F. Müller, 1785) + + Polyphemus pediculus (Linnaeus, 1758) + + + Scapholeberis mucronata (O.F. Müller, 1758) + Sida crystallina (O. F. Müller, 1776) + Simocephalus serrulatus (Koch, 1841)* + COPEPODA Acanthodiaptomus denticornis (Wierzejski, 1887) + + + + + + Megacyclops viridis (Jurine, 1820) + + + Thermocyclops dybowskii (Lande, 1890) + (+: present, *: new records for region, MC1970: Margaritora and Cottarelli 1970; A1999: Altındağ 1999; AY2000: Altındağ and Yiğit 2000, ÖM2017: Özdemir Mis et al. 2017) Discussion Cottarelli) and six Rotifera, (Dumont and De Ridder In freshwater ecosystems, zooplankton 1987), six Cladocera and two Copoepoda taxa were community have an important role in water quality described. All rotifer species were also found in this (Moss et al. 2003), and rotifers can also be used for study (Table 2). The most important difference this purpose (Gutkowska et al. 2013; Apaydın Yağcı between these two studies is the number of species et al. 2017). Rotifers are thought to become the found. The low number of zooplankton species, dominant taxa in many lakes over time (Wen et al. especially rotifers, maybe resulting from the 2011) and they can adapt to the degraded situation sampling procedures like using a broader mesh size, better than other similar taxa. According to Saksena taking one sample, etc. The cladoceran species, (1987), rotifers are the dominant group in freshwater Scapholeberis mucronata (O.F. Müller, 1758), ecosystems and as being compatible with this result, Daphnia hyalina Leydig 1860, Alona costata Sars, in our study, rotifer percentages (67%) were found to 1862, Acroperus harpae (Baird, 1843) and Alona be much higher than cladocerans (29%) and affinis (Leydig, 1860) were not found except P. copepods (4%), a situation similar to other truncatus. Mediterranean freshwater lakes (Saler 2017). K. Other detailed studies were carried out by longispina, K, cochlearis and K. quadrata known as Altındağ (1999) and Altındağ and Yiğit (2000). In cosmopolitan (Segers 2007) and they were observed these studies, rotifera phylum and zooplankton by all researchers. composition of Abant Lake were revealed. However, The first study on zooplankton community in in our study Lecane hamata (Stokes 1896), Colurella Abant Lake was carried out in 1970 (Margaritora and adriatica Ehrenberg, 1831, Synchaeta littoralis
Gürbüzer et al. 2019 - LimnoFish 5(1): 41-46 45
Rousselet, 1902 and Ascomorpha ecuadis were not Apaydın Yağcı M, Yeğen V, Yağcı A, Uysal R. 2017. A observed despite being present in Altındağ (1999) preliminary study on zooplankton species in different and also Alonella excisa (Fischer, 1854) and Sida aquatic habitats of Anatolia (Turkey). crystallina (O. F. Müller, 1776) were not observed Limnofish. 3(1): 45-50. despite being present in Altındağ and Yiğit (2000). doi: 10.17216/LimnoFish.277465 Atıcı T, Tokatlı C. 2014. Algal diversity and water quality Altındağ (1999) also pointed out that Asplanchna assessment with cluster analysis of four freshwater girodi is synonymous with A. brightwellii and we lakes (Mogan, Abant, Karagöl and Poyrazlar) of also described this taxon as A. girodi. Turkey. Wulfenia. 21(4):155-169. The most recent work regarding Abant Lake is of Çelekli A, Külköylüoğlu O. 2006. Net planktonic diatom a field study of Özdemir Mis et al., which were (Bacillariophyceae) composition of Lake Abant conducted in between 2002-2003 yet published in (Bolu). Turk J Bot. 30: 331-347. 2017. In this study, twenty-one taxa were determined De Smet WH. 1996. The Prolidae (Monogononta). and among them, 12 belonged to Rotifera, 6 to Amsterdam: SPB Academic Publishing, Vol. 4 102 p. Cladocera and 3 to Copepoda. Of these 21 taxa, 16 Dumont H, De Ridder M. 1987. Rotifers from Turkey. were mutual with our result (Table 2). A. denticornis Hydrobiologia. 147(1):65-73. Dügel M, Külköylüoğlu O, Kılıç M. 2008. Species has been found since 1970 in Abant Lake. assemblages and habitat preferences of Ostracoda Zooplankton differences between ours and this study (Crustacea) in Lake Abant (Bolu, Turkey). Belg. J. were Lecane luna and Platiyas quadricornis from Zool. 138 (1): 50-59. Rotifera, Alonella exigua from Cladocers and Erinç S, Bilgin T, Bener M. 1961. Abant Gölü’nün menşei Thermocyclops dybowskii from Copepoda fauna. hakkında. İ. Ü. Coğrafya Dergisi. 12: 184-187. Diversity of observed planktonic organisms can Filstrup CT, Downing JA. 2017. Relationship of be affected by the sampling method (for example chlorophyll to phosphours and nitrogen in nutrient- plankton net size and shape, etc.) therefore rich lakes, Inland Waters. 7(4): 385-400. zooplankton density and composition differences can doi: 10.1080/20442041.2017. 1375176 be due to the differences in sampling procedures Gannon EJ, Stemberger SR. 1978. Zooplankton as indicators of water quality. Trans Amer Micros Soc. (Gutkowska et al. 2012). Considering previous 97:16-35. studies, it is easy to conclude, species richness is doi: 10.2307/3225681 increased in the past decades especially for rotifers Gutkowska A, Paturej E, Kowalska E. 2012. Qualitative and cladoceran taxa. However, it is not clear whether and quantitative methods for sampling zooplankton in there is an increase in species richness, because the shallow coastal eustaries. Ecohydrology and methods used were not compatible with each other. Hydrobiology. 12(3):253-263. In this study, we believe high numbers of identified doi: 10.2478/v10104-012-0022-2 separate species compared to past stem from vertical Gutkowska A, Paturej E, Kowalska E. 2013. Rotifer and horizontal sampling in different habitats. We trophic indices as ecosystem indicators in brackish think that using standardized and commonly accepted coastal waters. Oceanol. 55 (4): 887-899. doi: 10.5697/oc.55-4.887 sampling methods will allow our predictions Gürbüzer P, Buyurgan Ö, Tekatlı Ç, Altındağ A. 2017. regarding future community compositions to be more Species diversity and community structure of accurate. zooplankton in three different types of water body within the Sakarya River Basin, Turkey. Turk J Zool. Aknowledgements 41(5): 848-859. A part of this article has been presented as an oral doi: 10.3906/zoo-1606-41 presentation in VIII. National Limnology Harding JP, Smith WA. 1974. A Key to the British Symposium and we thank three anonymous Freshwater Cyclopoid and Calanoid Copepods. 2nd reviewers for provided helpful comments on drafts of edition. Ambleside, UK: Freshwater Biological Association 54 p. this manuscript. Jeppesen E, Jensen P, Sondergaard M, Lauridsen T, Landkildehus F. 2000. Trophic structure, speceis References richness and biodiversity in Danish lakes: changes Akşıray F. 1959. Abant Gölü’nde sun’i ilkah yolu ile ilk along a phosphorus gradient. Freshwater Biol. alabalık üretilmesi hakkında. İ. Ü. Fen Fak. 45(2): 201-218. Hidrobiyoloji Dergisi, Seri A. 5(1-3):115-124. doi: 10.1046/j.1365-2427.2000.00675.x Altındağ A. (1999). A taxonomical study on the Rotifera Jeppesen E, Noges P, Davidson TA, Haberman J, Noges Fauna of Abant Lake (Bolu). Turk J Zool. T, Blank, K, Lauridsen TL, Sondergaard M, Sayer C, 23: 211-214. Laugaste R, Johansson LS, Bjerring R, Amsinck SL. Altındağ A, Yiğit S. (2000). Abant (Bolu) Gölü 2011. Zooplankton as indicators in lakes: a scientific- Zooplankton Faunasının Mevsimsel Değişimi. Ege J based plea for including zooplankton in the ecological Fish Aqua Sci. 17:(2) 9-18. quality assessment of lakes according to European
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LIMNOFISH-Journal of Limnology and Freshwater Fisheries Research 5(1): 47-59 (2019)
Fossil and Recent Distribution and Ecology of Ancient Asexual Ostracod Darwinula stevensoni (Ostracoda, Crustacea) in Turkey
Mehmet YAVUZATMACA * , Okan KÜLKÖYLÜOĞLU
Department of Biology, Faculty of Arts and Science, Bolu Abant İzzet Baysal University, Turkey
ABSTRACT ARTICLE INFO
In order to determine distribution, habitat and ecological preferences of RESEARCH ARTICLE Darwinula stevensoni, data gathered from 102 samples collected in Turkey between 2000 and 2017 was evaluated. A total of 1786 individuals of D. Received : 28.08.2018 stevensoni were reported from eight different aquatic habitats in 14 provinces in Revised : 21.10.2018 six of seven geographical regions of Turkey. Although there are plenty of samples Accepted : 30.10.2018 from Central Anatolia Region, recent form of the species was not encountered. Unlike recent, fossil forms of species were encountered in all geographic regions Published : 25.04.2019 except Southeastern Anatolia. The oldest fossil record in Turkey was reported from the Miocene period (ca 23 mya). Species occurred in all climatic seasons in DOI:10.17216/LimnoFish.455722 Turkey. D. stevensoni showed high optimum and tolerance levels to different ecological variables. Results showed a positive and negative significant * CORRESPONDING AUTHOR correlations of the species with pH (P<0.05) and elevation (P<0.01), respectively. [email protected] It seems that the ecological preferences of the species are much wider than Phone : +90 537 769 46 28 previously known. Our results suggest that if D. stevensoni is used to estimate past and present environmental conditions, attention and care should be paid on its ecology and distribution. Keywords: Ecologic preference and characterization, seasonality, stevensoni
Eski Aseksüel Ostrakod Darwinula stevensoni’nin (Ostracoda, Crustacea) Türkiye’deki Ekolojisi, Fosil ve Güncel Dağılımı Öz: Darwinula stevensoni’nin dağılımını, habitat ve ekolojik tercihlerini belirlemek için 2000 ve 2017 yılları arasında Türkiye’den toplanan 102 örnekten elde edilen veriler değerlendirilmiştir. Toplam 1786 D. stevensoni bireyi Türkiye’nin yedi coğrafi bölgesinin altısında bulunan 14 ildeki sekiz farklı sucul habitattan rapor edildi. İç Anadolu bölgesin de bol miktarda örnek olmasına rağmen, türün güncel formuna rastlanılmadı. Güncel formdan farklı olarak, türün fosil formu ile Güneydoğu Anadolu dışındaki tüm coğrafi bölgelerde karşılaşılmıştır. Türkiye’deki en eski fosil kayıt Miosen döneminden (yaklaşık 23 milyon sene önce) rapor edilmiştir. Türkiye’deki tüm mevsimlerde tür bulunmuştur. D. stevensoni farklı ekolojik değişkenlere yüksek optimum ve tolerans seviyeleri göstermektedir. Tür pH ile pozitif (P<0,05) fakat yükseklik ile negatif (P<0,01) anlamlı korelasyon göstermektedir. Görülmektedir ki türün ekolojik tercihleri daha önce bilinenden daha geniştir. Sonuçlar, D. stevensoni’nin güncel ve geçmiş çevre koşullarını tahmin etmek için kullanılması halin de ekolojisine ve dağılımına dikkat edilmesi gerektiğini göstermektedir. Anahtar kelimeler: Ekolojik tercih ve karakterizasyon, mevsimsellik, stevensoni
How to Cite Yavuzatmaca M, Külköylüoğlu O, 2019. Fossil and Recent Distribution and Ecology of Ancient Asexual Ostracod Darwinula stevensoni (Ostracoda, Crustacea) in Turkey. LimnoFish. 5(1): 47-59. doi: 10.17216/LimnoFish.455722
Introduction (Benson 1990; Külköylüoğlu 1999). Also, because of Ostracods are small (0.3-5 mm long), the easily fossilization of calcium carbonated bivalved (carapaces) aquatic creatures that are carapaces, they are commonly used in widely distributed in a variety of marine biostratigraphy, paleobiology, paleoclimatology, and non-marine environments (Meisch 2000). paleolimnology and paleoecology studies (Ruiz et al. They show species-specific responses to the changes 2013). In this sense, the autecology of individual in different ecological conditions; therefore, species has an important role since ecology of they can be used as bioindicator species to recent species help paleontologists to widen estimate possible environmental deterioration their perceptions to understand types of
48 Yavuzatmaca and Külköylüoğlu 2019 - LimnoFish 5(1): 47-59 paleoenvironmental conditions based on fossil distribution of species (cf. Figure 2 in this paper). ostracods (Carbonel et al. 1988). Species has been collected in lotic, lentic and The genus Darwinula is the type genus of the interstitial habitats and it is ecologically family Darwinulidae that includes only the type characterized as thermoeuryplastic, oligorheophilic, species Darwinula stevensoni. Because of the titanoeuryplastic and mesohalophilic (Meisch 2000). absence of males in fossils (since Mesozoic) and in Until now, the ecology of D. stevensoni has not been living populations, superfamily Darwinuloidea have widely evaluated except some papers partially survived asexually over 200 million years (Martens discussed its status in local and/or regional et al. 2003). Similar to bdelloid rotifers (Mark Welch perspectives (e.g., Ranta 1979; Rossetti and Martens and Meselson 2000; Mark Welch et al. 2004) and 1996; Gandolfi et al. 2001a, 2001b; Van Doninck et oribatid mites (Maraun et al. 2004), darwinuloid al. 2003a; Rossi et al. 2002, 2004; Higuti et al. 2009a; ostracods (Schön and Martens 2003) have also been Van den Broecke et al. 2013). Therefore, the aims of suggested as one of the “putative ancient asexual” the present study are to (i) determine geographic and groups in animal kingdom. However, although its local distribution of both fossil and living populations persisting asexuality is known, a few reports of rare of the species among different aquatic habitats, (ii) males of the species by Turner (1895) and Brady and estimate ecological preferences of D. stevensoni in Robertson (1870) and one male of Vestalenula Turkey, and (iii) evaluate species ecological cornelia (Smith et al. 2006) has also been questioned tolerance and optimum ranges for those of particular within the family. Subsequently, Martens and Schön environmental variables. (2008) indicated D. stevensoni as a strong candidate in darwinuloids to being a true ancient asexual. In support of this view, Schön et al. (2009) stated that occasional males may be produced in many asexual species because of mutations in the regulatory cascade controlling sex and the males produced are not functional. The presence of female fossils dating back to 25 million years (Straub 1952) and the absence of functional or atavistic males in recent and fossil species (Schön et al. 2009) enforce the ancient Figure 1. Darwinula stevensoni. Left valve external view (Scale bar 100 µm). asexuality of D. stevensoni. D. stevensoni is a small sized (0.6-0.8 mm in Materials and Methods length) (Meisch 2000) ostracods. Carapace of species is characteristically cigar shaped with unequal valves Study sites and Sampling (Figure 1). Right valve extends left valve on all sides A total of 102 samples from 14 provinces (72 except hinge. The posterior margin of carapace is samples from Bolu, 1 in Gaziantep, 2 in Ordu, 3 in wider than anterior. The widening of posterior part is Adıyaman, 3 in Burdur, 2 in Hatay, 3 in Mardin, 2 in due to the developing of a brood chamber since Muş, 4 in Kütahya, 1 in Mersin, 2 in Sakarya, 2 in species is viviparous parthenogenetic unlike most of Isparta, 3 in Antalya and 2 in Muğla) of Turkey were other ostracods (Cypridoidea and Cytheroidea) collected between the years 2000 and 2017 (Figure 2, (Rossetti and Martens 1996). Generally, species Appendix). All the measurements were taken in situ obtain about ten eggs in this brood chamber, but this before ostracods were collected to prevent the mixing number can be changed up to 13 (Külköylüoğlu pers. of water and to obtain the actual values of variables. obs.). Later, juveniles may have more than one Ostracod samples were collected with a standard molting stage within brood space until hatched. sized (200 µm) d-frame hand net and fixed in 70% Muscle scars that control the opening and closing of ethanol. valves arranged in a characteristic circular rosette Physico-chemical variables (pH, dissolved shape, which includes 9 - 12 spots. Additionally, oxygen (DO, mg/L), percent oxygen saturation species do not swim because of the absence of (%DO), water temperature (Tw, °C), electrical natatory setae on the second antennae of species and conductivity (EC, µS/cm), salinity (Sal. ‰), total so it is a typical benthic form (for more taxonomic dissolved solids (TDS, mg/L) and redox potential remarks see Rossetti and Martens 1996; Meisch (ORP, mV)) were measured by a YSI-85 model of 2000). oxygen-temperature and HI-98150 pH-ORP meter D. stevensoni showed a cosmopolitan distribution from sapling sites in Bolu and Ordu where (Meisch 2000) except from Antarctic region, Pacific geographic data (coordinates and elevation) was region and Oceonic Islands (Martens et al. 2013). recorded with a Garmin GPS-12XL. In Gaziantep Van Doninck et al. (2003a) presented the global and Hatay provinces, Tw, pH, EC and salinity values
Yavuzatmaca and Külköylüoğlu 2019 - LimnoFish 5(1):47-59 49 were obtained with a Delta OHM pH/conductivity for the rest of the provinces were done by YSI meter while air temperature was recorded with a Professional Plus and Testo 410-2 anemometer and a Testo 410-2 anemometer, and coordinates and Garmin etrex Vista H GPS (for elevation and elevation with a Garmin GPS-45. The measurements coordinates).
Figure 2. Distribution of living D. stevensoni in 27 provinces of Turkey. * indicates provinces (in bold) sampled during the current study, + represents the species previously reported in the literature. *+ displays the species records in this study and previously as well. Citations for the previous reports; İstanbul (Külköylüoğlu pers. obs. unpublished data), Kocaeli (Akdemir pers. obs. unpublished data), Bursa (Altınsaçlı and Griffiths 2001a); Balıkesir (Altınsaçlı and Griffiths 2001b); Düzce (Gülen 1985); Afyon (Gülen 1985); Denizli (Altınsaçlı and Mezquita 2008); İzmir (Meriç et al. 2010); Isparta (Özuluğ et al. 2001); Konya (Akdemir 2004); Aksaray (Altınsaçlı 2004); Şanlıurfa (Özuluğ and Dökümcü 2014); Diyarbakır (Gülen et al. 1996; Akdemir and Külköylüoğlu 2011); Erzincan (Akdemir and Külköylüoğlu 2014). In the laboratory, samples were filtered Statistical Analyses through four standardized sieves (0.5, 1.0, 1.5 and 2.0 The tolerance (tk) and optimum (µk) estimates of mm mesh size) under tap water and then specimens the species for different ecological variables were were separated from sediment under calculated by using C2 software after using a transfer stereomicroscope and fixed in 70% ethanol function of weighted averaging regression (Juggins for further studies. Taxonomic identification 2003). A non-parametric Spearmen Rank Correlation was done according to the carapace and soft body was applied to see the the levels of correlations parts dissected in lactophenol solution by using between species and different variables (IBM-SPSS taxonomic key of Meisch (2000) under Statistics version 21). a light microscope (Olympus BX-51). According to Meisch (2000), D. stevensoni was ecologically Results characterized for salinity limnetic (freshwater) range We encountered 1786 individuals of (<0.5 ‰), oligohaline (0.5-5 ‰), mesohaline (5-18 D. stevensoni from eight different aquatic habitats ‰), polyhaline (18-30 ‰), euhaline (30-40 ‰) and in 14 provinces (Figure 3). These provinces were hyperhaline (≥40 ‰) and water temperature (cold found in six geographical regions of Turkey stenothermal, oligothermophilic, mesothermophilic except Central Anatolia Region (Figure 2). The (between two previous), polythermophilic highest (1117) and lowest (1) individual numbers and warmstenothermal). On the other hand, were reported in dams and pond, respectively. because this ecological characterization does not Although the number of lakes and springs sampled classify water temperature ranges for were approximately 9 and 6 times more than the freshwater habitats, we followed the offering of Chu number of troughs, respectively, the number of et al. (2009) and Olivero-Sheldon et al. (2014). The individuals found in troughs (277) are more than ranges as very cold (<12.8 °C), cold (<18 °C), cold the number of individuals in both habitats (234) cool (>18 at <21 °C) and warm (>21 °C) were used. (Figure 3). In general, D. stevensoni was mostly All the specimens were kept in Limnology encountered in May, December, and January months Laboratory of Bolu Abant İzzet Baysal University, of the seasons (spring and winter) when it was Turkey. collected in all months of summer and autumn.
50 Yavuzatmaca and Külköylüoğlu 2019 - LimnoFish 5(1): 47-59
Accordingly, we found the species from all the four The optimum and estimated tolerance levels of D. seasons in Turkey. The distribution of fossil forms stevensoni for eight different variables are given in of species throughout Turkey were given in the Table 1. D. stevensoni showed positive and negative Figure 4. Accordingly, fossil records indicated significant correlations with pH (r = 0.218, N = 82, that the species has been known from Miocene (about P<0.05) and elevation (r = -0.280, N = 101, P<0.01), 25 mya) in Turkey. respectively.
Figure 3. Sampling numbers (NuSmp) of eight different habitats and individual numbers of D. stevensoni (NuInd) in these habitats Table 1. The optimum (µk) and estimated tolerance (tk) levels of D. stevensoni for pH, dissolved oxygen (DO), electrical conductivity (EC), water temperature (Tw), oxidation and reduction potential (ORP), elevation (Elev), salinity (Sal) and total dissolved solids (TDS). Abbreviations: Count, the number of sampling sites where species found; Max, the maximum number of individuals of concerned species among sampling sites and N2, Hill’s coefficient value that indicates the measure of effective number of occurrences.
pH DO EC Tw
µk tk µk tk µk tk µk tk Count Max N2
101 214 17.38 6.82 3.37 9.17 3.68 351.62 168.67 19.44 6.22
ORP Elev Sal TDS µk tk µk tk µk tk µk tk Count Max N2
101 214 17.38 114.92 82.56 708.62 232.33 0.13 0.07 142.00 104.31
Darwinula stevensoni
The minimum and maximum values of Discussion pH (6.90-10.60), DO (0.32-18.31 mg/L), Along with the results of the present study and DO% (3.30-171.50 %), EC (21-844 µS/cm), Tw literature, recent living forms of the species has been (6.10-31 °C), Ta (13-40.20 °C), ORP (-107.27- now recorded from 27 provinces located in all seven 240.60 mV), Elev (39-2163 m a.s.l.), Sal. (0-0.42 ‰) geographic regions of Turkey (Figure 2). On the and TDS (0.06-503.17 mg/L) of sampling sites where other hand, the fossil forms of the species D. stevensoni collected. Accordingly, the species D. stevensoni were only reported from 20 provinces lives in waters with fresh to slightly brackish water covering six geographic regions of Turkey except ranges and it is characterized as a meso- Southeastern Anatolia. More than half of these polythermophilic. 20 provinces were located in the western parts of
Yavuzatmaca and Külköylüoğlu 2019 - LimnoFish 5(1):47-59 51
Turkey (Figure 4). Of these provinces, in Sakarya, with ostracods in other parts (north, south, and east) dead specimens of D. stevensoni were taken from of Turkey. As a result, D. stevensoni (in fossil and superficial sediments at 18 and 6 m depth of Lake recent forms) has been reported in 44.4% of 81 Sapanca by Nazik et al. (2011) but they did not provinces, Turkey. Among these provinces, specify the age of these sediments. Meisch (2000) eleven have fossil and recent forms and of the rest, 16 indicated that fossil record is known from Mid- and nine provinces have only recent and fossil Oligocene (ca 28 mya) to present but oldest fossil forms, respectively (Figures 1 and 3). Accordingly, record in Turkey has been reported from the Miocene with the current study, geographical distribution of period (ca 23 mya) (see Figure 4). These data actually D. stevensoni has been now expanded throughout indicate the lack of paleontological studies dealing Turkey.
Figure 4. Fossil reports of D. stevensoni from Kırklareli (Witt 2011), İstanbul (Şafak et al. 1999; Meriç et al. 2000; Şekeryapan 2011), Yalova (Rückert-Ülkümen and Yiğitbaş 2007), Kocaeli (Matzke-Karasz and Witt 2005; Rückert- Ülkümen et al. 2006; Schneider et al. 2005), Sakarya (Nazik et al. 2011), Bursa (Freels 1980; Franz et al. 2006; Meriç et al. 2009; Nazik et al. 2011), Çanakkale (Atay and Tunoğlu 2002), Afyon (Demirer et al. 2017), Manisa (Witt 2003), Aydın (Tuncer and Tunoğlu 2015), Denizli (Gökçen 1979a, 1979b; Freels 1980; Şafak 2010), Muğla (Gökçen 1979a, 1979b), Burdur (Freels 1980), Adana (Nazik et al. 1992, 1999), Kahramanmaraş (Freels 1980), Konya (Freels 1980), Niğde (Nazik and Gökçen 1989), Erzincan (Freels 1980), Erzurum (Freels 1980) and Bolu (Tunoğlu et al. 2012) provinces in Turkey. Abbreviations: Early (E), Middle (M), Late (L), Lower (Lo), Upper (U), Miocene (Mi), Pannonian (Pan), Pontian (Po), Pliocene (P), Pleistocene (Ple), and Holocene (H). Darwiula stevensoni prefers ponds, lakes and Additionally, species was commonly found in slow flowing streams (Meisch 2000). Szlauer- dam, lake, and springs (Figure 3). These habitats are Łukaszewska (2014) and Ruiz et al. (2013) suggested generally permanent almost throughout the year. This D. stevensoni as a lake littoral species and benthos of confirms the suggestions of Palacios-Fest (2002- shallow aquatic bodies, respectively. Pérez et al. 2003) as the occurrence of the species might be (2011) and Marchegiano et al. (2017) indicated its associated with long term water permanence. negative correlation with depth. Contrary to common In addition, Escrivà et al. (2014) emphasized that belief that species is generally found in shallow D. stevensoni was one of the most common species aquatic bodies, it has been reported from 0 to a in reservoirs. Until now, it has been reported from maximum 20 m depth in literature (Meisch 2000; variety of habitats, such as ponds, slow flowing Pérez et al. 2010; Lorenschat and Schwalb 2013; streams, lakes, springs, rivers, cenotes, troughs, Lorenschat et al. 2014) but species had a low coastal lagoons, wetlands (slough), artificial dam optimum (2.3 m) and tolerance (3.8 m) levels to depth lakes, marshes, interstitial ground water, hot springs, (Lorenschat and Schwalb 2013). The occurrence of reed beds, rice field, peat bogs, ditch and canal species in the water bodies with a maximum 1 m (Gülen 1985; Mezquita et al. 1999a, 1999b; depth herein supports the common occurrence of Meisch 2000; Laprida et al. 2006; Higuti et al. 2009b; species in shallow waters although it has been Pieri et al. 2009; Pérez et al. 2011; Akdemir and encountered in a maximum of 20 m depth. Külköylüoğlu 2014; Escrivá et al. 2014; Mazzini et
52 Yavuzatmaca and Külköylüoğlu 2019 - LimnoFish 5(1): 47-59 al. 2014; the current study). Based on the data observed the presences of 15 juveniles in brood available, one may consider the fact that species cavity. Accordingly, the species seems to have seems to have high levels of plasticity for habitats characteristics of K-selected or r-K continuum as proposed by Ranta (1979). As seen in Figure 3, species (Van Doninck et al. 2003b). Furthermore, our results also reinforced the plasticity of species when comparing Darwinulids with other ostracods, for habitats. This means that there are no specific they generally have low fecundity (Geiger 1998; Van habitat preferences of species, which may present den Broecke et al. 2013) and produce less eggs (0.02- certain conditions. However, highest numbers 0.07 layed eggs per day (Gandolfi et al. 2001b) and of individuals per site were reported from an artificial maximum 20 eggs per generation (Ranta 1979) that trough (92) that is followed by slough (63) and lower the number of cell division and thus the dam (27) (Figure 3). These habitats are sensitive to mutation rate falls in D. stevensoni (Van Doninck et the outside effects of anthropogenic, seasonal and al. 2003b). climatic changes (Külköylüoğlu et al. 2013; Uçak et Martens and Tudorance (1991) recorded the al. 2014). Besides to habitat plasticity of the species, escape of species from the places with high it has been collected from the habitats with sapropel temperature values in a tropical Ethiopian lake. (mud), sand, rocks, stones and gravel sediment types Indeed, this observation was actually supported by (Altınsaçlı and Griffiths 2001b; Szlauer- the studies of Van Donnick et al. (2003a) that the Łukaszewska and Kowaluk-Jagielska 2011; species survival was shown to decrease with Lorenschat and Schwalb 2013) that reinforce the increasing temperature. Besides, Pérez et al. (2011) proposal of Ranta (1979). In addition, Külköylüoğlu reporting a negative correlation between and Vinyard (2000) reported it from muddy and D. stevensoni and temperature confirmed the sandy sediments with high dissolved oxygen previous observation. When we look at the optimum concentration. and tolerance values of the species for water The reproduction periods of the species show temperature 16.4-1.2 °C (Mezquita et al. 2005), slightly different patterns. For example, reproduction 20.6-5.3 °C (Lorenschat and Schwalb 2013) and of the species takes from May to October 19.44-6.22 °C (this study) (Table 1) it appears that (Meisch 2000) while it takes from March to species can tolerate a broad temperature range from September in a temperate pond in Belgium cold to warm waters. Along with these information, (Van Doninck et al. 2003b). These previous authors wide temperature ranges of species from 4 were also reported species from January to (in subarctic) to 35 °C (Van Doninck et al. 2003b; November in this temperate pond. Besides, Külköylüoğlu 2013) support the suggestion of Külköylüoğlu (1999) collected species from Gandolfi et al. (2001b) and Anàdon et al. (2012) February to November from springs of Nevada. who characterized the species as eurythermal Martín-Rubio et al. (2005) reported species from (tolerating and adapting to wide range of Lake Caicedo de Yuso-Arreo (Spain) in January, temperature) and thermoeuryplastic (a wide range of February, March, April, June - August, temperature tolerance), respectively. and November when Scharf and Viehberg (2014) When we look at the literature for the species encountered species in February, April, June, July, occurrence patterns in different areas, it appears that September and October in Germany. The occurrence its occurrence was reported to be positively of the species in April from Lake Meyil (Konya, related to biological oxygen demand (BOD), Turkey) (Akdemir 2008) and from May to January ammonium content (Mezquita et al. 1999a), DO, herein is now supported previously reported pH (Martens and Tudorance 1991), low (Rieradevall reproduction period of species. The occurrence of and Roca 1995) or highest water temperatures species in all climatic seasons and almost in all (Escrivá et al. 2014), warm water, carbonated water months in Turkey supports the founding of species rich with sulfate and chloride (Mezquita et al. 1999b), throughout the year (Hiller 1972; Altınsaçlı and but the relationship with iron content was negative Griffiths 2001a). Martens and Tudorance (1991) (Iglikowska and Namiotko 2012). The result of the also pinpointed that D. stevensoni is a perennial previous studies supports the suggestion of species in a tropical Ethiopian lake. Therefore, Külköylüoğlu and Vinyard (2000) as D. stevensoni D. stevensoni is showed as a eurychronal species. prefers less saline waters. Similarly, Van Doninck et All the data provided in here enforces its life cycle al. (2003a) suggested that survival of D. stevensoni is with about one or more years, during which two or declined with increasing in salinity. Besides, Pérez et more generations are produced. In each generation, al. (2011) reported the species tolerating electrical females can carry maximum 11-12 embryos within conductivity optima at <700 µS/cm when Mezquita their brood pouch (Van Doninck et al. 2003b; et al. (2005) and Lorenschat and Schwalb (2013) Gandolfi et al. 2001b). However, Horne et al. (1998) announced the optimum and tolerance level of
Yavuzatmaca and Külköylüoğlu 2019 - LimnoFish 5(1):47-59 53 species for electrical conductivity as followings Ranta (1979) delineated that D. stevensoni 3.09 ± 0.39 mS/cm and 239.1 ± 35.3 µS/cm (for prefers highly oxygenated waters to aerate its eggs in salinity 0-0.04 ‰), respectively. The low optimum brood chamber as stated by Külköylüoğlu and and tolerance levels of species for EC Vinyard (2000) and Rossetti et al. (2004). The high (351.62 ± 168.67) and salinity (0.13 ± 0.07) optimum and tolerance of species for dissolved herein (Table 1) strengthened these previous oxygen 8.4 ± 2.1 mg/L in Mezquita et al. (2005), 7.9 findings. On the other hand, Martens (1990) noted ± 3.1 mg/L in Lorenschat and Schwalb (2013) and the presence of species in the East African Lake Shala herein (see Table 1) support the proposal of Ranta with 16–21 g/L salinity. Recently, Mischke et al. (1979). On the other hand, Escrivà et al. (2014) (2014) indicated the overcoming of species to large proposed preferences of species for lowest dissolved specific conductivity fluctuation but suggesting low oxygen. Although species might die under oxygen optimum and tolerance of the species in 3.164 µS/cm depletion (Rieradevall and Roca 1995), species can and 0.916 µS/cm of EC values. The minimum and live over one month (38 days) under hypoxic maximum value of electrical conductivity 21–9600 conditions (ca. 0.12 ml/L oxygen) in laboratory (after 15 days 100 % mortality observe) μS/cm (the conditions (Rossi et al. 2002). The minimum and current study; Gandolfi et al. 2001a) and salinity maximum DO ranges of the species (0.32-18.31 range 0-15 ‰ (the current study; Meisch 2000) for mg/L) herein indicate that species may tolerate from the species in literature strengthen the tolerating low to high oxygen concentrations. Moreover, there ability of species to salinity and EC fluctuations. is no any studies dealing the number or quality of Consequently, all of them fortify the euryhaline eggs of species in low and high DO concentration and characteristics of species (Gandolfi et al. 2001b) and so the proposal of Ranta (1979) may be acceptable the presence of species from distilled water to sea until otherwise stated. In addition, minimum and water (Van Doninck et al. 2003a). maximum values of ambient air temperature (12- The positive correlation of D. stevensoni with pH 40.20 ºC) (Horne 2007; the current study), calcium herein endorses the recommendation of Rossetti et al. (5.25-80.80 mg/L) (Higuti et al. 2009a; Pérez et al. (2004). They found a close association between 2015) and magnesium (2.30-100.80 mg/L) (Holmes species and pH in eutrophic freshwater wetlands of 1997; Pérez et al. 2015) contents of water bodies northern Italy. In contrast, negative correlation was where species collected. recorded between the species and pH (Pérez et al. The above-mentioned information and wide 2011; Marchegiano et al. 2017). However, the high range of environmental variables for species confirm optimum and tolerance levels of species for pH as the presence of all characteristics of the idea called following 7.74 ± 0.40 (Mezquita et al. 2005) and “general purpose genotype” (GPG) in D. stevensoni 8.62 ± 0.26 (Lorenschat and Schwalb 2013) were (Rossi et al. 2002; Van Doninck et al. 2002). GPG announced from several different aquatic bodies. emphasizes the production of different phenotypes by When we compile all data and compare optimum and a genotype across a wide range of environmental tolerance (6.82 ± 3.37, Table 1) levels of the species conditions that allow species survive with high along with min/max values (5.5–10.60) (Ruiz et al. fitness in a wide range of habitats (Baker 1965; 2013; the current study) for pH, it can be clearly seen Geiger et al. 1998). This character of D. stevensoni that the D. stevensoni is of wide ranges of pH reinforce the idea of Vandel (1928) as that tolerance. “parthenogenetic (i.e., ancient asexual D. stevensoni Additionally, species was negatively correlated herein) forms can be found in much wider areas” and with elevation in the current study. Possible effects of referring to its long living without sex. Accordingly, elevation on the physico-chemical characteristics of Külköylüoğlu (2013) called the species as the aquatic bodies are widely discussed (Reeves et al. “cosmoecious species” to distinguish it from other 2007; Rogora et al. 2008). Accordingly, it seems that species because of its wide geographical distribution elevation can be effective on the abundance but its and with relatively wide ecological tolerance ranges effect may not be significant on the occurrence in variety of aquatic habitats. This view implies to and distribution of species at high elevations. This is take attention of scientists who want to use because it has been reported from sea level D. stevensoni as a potential bioindicator species to (Külköylüoğlu pers. obs.) to 4000 m a.s.l. estimate past conditions and to determine water (Laprida et al. 2006). Indeed, generally individual quality values of the present habitats. Additionally, numbers larger than 100 were found from species is ecologically characterized as stated by the sampling sites between 700-900 m a.s.l. and at Meisch (2000), thermoeuryplastic and it encountered lowest elevation when the range is 39 (122 from freshwater range to mesohaline range. individuals) – 2163 (5 individuals) m a.s.l in the As mentioned above, fossils of the species are present study. known from Miocene and distribution of fossils
54 Yavuzatmaca and Külköylüoğlu 2019 - LimnoFish 5(1): 47-59 forms are mostly known from western parts of doi: 10.1080/00222930701851735 Turkey. Indeed, occurrences of both living and fossil Anadón P, Gliozzi E, Mazzini I. 2012. Geochemical forms partially overlap in some regions, indicating and palaeoecological analyses of Mid Pleistocene the long lasting surviving possibilities of the species to Holocene astracod assemblages from in these regions. On the other hand, considering the Valle di Castiglione (Italy): Palaeoenvironmental and palaeoclimatic assessment. Dev Quat Sci. 17:217- fact that the species has not been found from 240. hundreds of recent and paleontological samples, doi: 10.1016/B978-0-444-53636-5.00013-5 we may assume that D. stevensoni has not been able Atay G, Tunoğlu C. 2002. Ostracoda fauna and their to reach to these regions. We believe that absence of bioprovince of the Kilitbahir drilling samples the species from these samples may also be related to (Eceabat/Çanakkale). Yerbilimleri (Bulletin of Earth several other a/biotic factors but it is also possible Sciences Application and Research Centre of that this is just a matter of time. Besides, as seen from Hacettepe University). 26:119-144. Figures 1 and 2, as much as contemporary studies on Baker HG. 1965. Characteristics and modes of origins of living recent forms, paleontological studies are far weeds. In: Baker HG, Stebbins GL, editors. The away from understanding of their distribution in Genetics of Colonizing Species. New York, NY, USA: Academic Press. p. 147-172. Turkey (if not the whole world). Thus, our study Benson RH. 1990. Ostracoda and the discovery of global strongly suggests the need for future studies not only Cainozoic palaeoceanographical events. In: Whatley to understand for the distributional patterns of R, Maybury C, editors. 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Appendix Sampling numbers (SpNu), name of the county, habitat type, date and number of individuals of D. stevensoni (Abundance). na means not available. Habitat types: 1, Lake; 2, Creek; 3, Trough; 4, Dam; 5, Stream; 6, Pond; 7, Spring and 8, Slough.
County Habitat County Habitat SpNu Date Abundance SpNu Date Abundance name type name type 1 Bolu 4 30.06.2000 2 52 Bolu 1 26.10.2003 1 2 Bolu 4 28.07.2000 1 53 Bolu 1 26.10.2003 1 3 Bolu 4 28.07.2000 3 54 Bolu 1 31.10.2003 1 4 Bolu 4 31.08.2000 3 55 Bolu 1 31.10.2003 2 5 Bolu 4 29.09.2000 2 56 Bolu 1 31.10.2003 3 6 Bolu 4 29.09.2000 2 57 Bolu 7 15.11.2003 6 7 Bolu 4 29.09.2000 3 58 Bolu 1 30.11.2003 2 8 Bolu 4 29.09.2000 4 59 Bolu 7 13.12.2003 1 9 Bolu 4 29.09.2000 4 60 Bolu 7 17.01.2004 6 10 Bolu 4 29.09.2000 8 61 Bolu 1 29.05.2004 1 11 Bolu 4 29.09.2000 34 62 Bolu 1 29.05.2004 1 12 Bolu 4 29.10.2000 2 63 Bolu 1 28.07.2004 1 13 Bolu 4 29.10.2000 2 64 Bolu 1 28.07.2004 1 14 Bolu 4 29.10.2000 3 65 Bolu 7 27.08.2004 1 15 Bolu 4 29.10.2000 3 66 Bolu 7 27.08.2004 2 16 Bolu 4 29.10.2000 3 67 Bolu 1 30.08.2004 1 17 Bolu 4 29.10.2000 28 68 Bolu 7 18.09.2004 5 18 Bolu 4 29.10.2000 201 69 Bolu 1 25.09.2004 2 19 Bolu 4 26.11.2000 32 70 Bolu 7 17.10.2004 2 20 Bolu 4 31.12.2000 1 71 Bolu 7 13.11.2004 1 21 Bolu 7 31.05.2001 10 72 Ordu 3 15.06.2010 1 22 Bolu 4 30.06.2001 50 73 Ordu 1 15.06.2010 77 23 Bolu 4 26.07.2001 2 74 Gaziantep 7 21.07.2010 9 24 Bolu 4 26.07.2001 6 75 Adıyaman 7 17.07.2012 52 25 Bolu 4 26.07.2001 10 76 Adıyaman 6 18.07.2012 1 26 Bolu 4 26.07.2001 14 77 Adıyaman 4 19.07.2012 2 27 Bolu 4 26.07.2001 22 78 Hatay 2 01.08.2012 5 28 Bolu 4 26.07.2001 32 79 Hatay 2 06.08.2012 2 29 Bolu 4 26.07.2001 101 80 Burdur 7 31.08.2012 1 30 Bolu 7 28.08.2001 1 81 Mardin 2 14.08.2013 1 31 Bolu 4 28.08.2001 19 82 Mardin 3 14.08.2013 62 32 Bolu 4 28.08.2001 101 83 Mardin 5 15.08.2013 3 33 Bolu 4 30.09.2001 3 84 Muş 7 18.08.2013 1 34 Bolu 4 30.09.2001 101 85 Muş 1 19.08.2013 5 35 Bolu 4 30.09.2001 101 86 Sakarya 7 10.05.2014 1 36 Bolu 1 6-7.10.2001 1 87 Sakarya 8 10.05.2014 122 37 Bolu 1 13.10.2001 1 88 Kütahya 4 21.09.2014 3 38 Bolu 1 13.10.2001 3 89 Kütahya 8 21.09.2014 4 39 Bolu 1 14.10.2001 1 90 Kütahya 2 21.09.2014 9 40 Bolu 4 30.10.2001 1 91 Kütahya 3 21.09.2014 214 41 Bolu 4 30.10.2001 101 92 Mersin 7 06.10.2015 3 42 Bolu 4 30.10.2001 101 93 Antalya 2 17.08.2017 6 43 Bolu 1 11.11.2001 1 94 Isparta 1 18.08.2017 2 44 Bolu 4 30.11.2001 1 95 Antalya 2 19.08.2017 2 45 Bolu 4 30.11.2001 2 96 Burdur 1 21.08.2017 3 46 Bolu 4 30.11.2001 3 97 Muğla 1 22.08.2017 2 47 Bolu 1 31.05.2002 1 98 Muğla 2 25.08.2017 1 48 Bolu 1 31.05.2002 3 99 Antalya 2 13.10.2017 1 49 Bolu 1 30.07.2002 1 100 Isparta 1 16.10.2017 2 50 Bolu 1 29.08.2003 8 101 Burdur 1 19.10.2017 3 51 Bolu 1 31.08.2003 1 102 Bolu 7 na 1
LIMNOFISH-Journal of Limnology and Freshwater Fisheries Research 5(1): 60-69 (2019)
Çayköy Deresi (Eğirdir-Isparta)’ndeki Eğirdir Siraz (Capoeta pestai Pietschmann, 1933)’larının Helmint Parazitleri Üzerine Bir Araştırma
Nesrin EMRE1* , Ayşegül KUBİLAY 2
1Akdeniz Üniversitesi, Eğitim Fakültesi Fen Bilgisi Anabilim Dalı, 07058, Antalya 2 Isparta Uygulamalı Bilimler Üniversitesi Eğirdir Su Ürünleri Fakültesi, Hastalıklar Anabilim Dalı, 32260, Isparta
ÖZ MAKALE BİLGİSİ
Bu çalışmada, Çayköy deresi (Eğirdir-Isparta)’nden elektroşokerle avlanan ARAŞTIRMA MAKALESİ Capoeta pestai (Pietschmann, 1933)’nin helmint parazitleri araştırılmıştır. Araştırma süresince toplam 150 birey incelenmiştir. Çalışma sonucunda konak Geliş : 12.09.2018 balıkta Platyhelminthes grubundan bir tür digenean (Allocreadium isoporum Düzeltme : 12.10.2018 Loos, 1894) ve Nemathelminthes grubundan bir tür nematod (Rhapdochona Kabul : 19.10.2018 denudata, Dujardin, 1845) bulunmuştur. Her mevsimde örneklenen bireylerden toplam 188 A. isoporum ve 592 adet R. denudata olmak üzere toplam 780 adet Yayım : 25.04.2019 parazit tespit edilmiştir. Her iki parazitin de mevsim, cinsiyet, boy ve yaş değişkenlerine göre; yaygınlık, ortalama yoğunluk, bolluk ve toplam parazit DOI:10.17216/LimnoFish.459292 sayıları belirlenmiştir. * SORUMLU YAZAR Anahtar kelimeler: Çayköy Deresi, Capoeta pestai, helmint parazitler [email protected] Tel : 0505 477 54 46 A Research on Helminth Parasites of Capoeta pestai (Pietschmann, 1933) from Çayköy Stream (Eğirdir-Isparta) Abstract: In this study, the helminth parasites of Capoeta pestai (Pietschmann, 1933) that was caught by electro shocker in Çayköy stream (Eğirdir-Isparta) has been investigated. Throughout study, 150 individuals have been taken into consideration. As a result, digenean belongs to Platyhelminthes group and nematod belongs to Nemathelminthes have been determined. 188 A. isoporum Loos, 1894 and 592 R. denudata, Dujardin,1845 have been obtained from individuals that was sampled in every season. Mean intensity, total number, abundance and prevalence of both parasites has been set in regard to season, host size, age and sexes. Keywords: Çayköy Stream, Capoeta pestai, helminth parasite
Alıntılama Emre N, Kubilay A, 2019. Çayköy Deresi (Eğirdir-Isparta)’ndeki Eğirdir Siraz (Capoeta pestai Pietschmann, 1933)’larının Helmint Parazitleri Üzerine Bir Araştırma. LimnoFish. 5(1): 60-69. doi: 10.17216/LimnoFish.459292
Giriş Doğal yaşam biçiminden koparılan bu türlerin gerek Su ürünleri içinde balık gerek sayısal ve gerekse beslenme ve gerekse sağlık anlamında problemleri ekonomik yönden önemli bir yer tutmaktadır. olmakta ve bununla ilgili çözümler için çalışmalar Ülkemizdeki tatlı sularda yaşayan balık tür sayısı da yapılmaktadır. Sucul ekosistemlerde besin zincirinin son yıllarda yapılan revizyonlarla 27 familyaya ait son basamaklarında yer alan ve insanlar için önemli 92 cins ve 371 türe ulaşmıştır. Bu türlerin bir protein kaynağı olan balıklar, farklı parazit yarısından fazlasının Cyprinidae familyası üyesi türlerine konaklık ederler. Bu parazitler ise balıklarda oldukları saptanmıştır. Bunların 19’unu ciddi hastalıklara ve ekonomik kayıplara neden Capoeta cinsine ait türler oluşturmaktadır (Kuru vd. olabilirler. Parazitler doğal balık populasyonlarında 2014). az düzeyde görüntü zararlarına neden olduğu gibi, Gerek ülkemizde ve gerekse dünya balık yetiştirilen balıklarda önemli hastalıkların üretiminde yetiştiricilikle avcılık arasındaki istihsal oluşmasına zemin hazırlayabilir; patolojik oranları oldukça biribirine yaklaşmıştır (Anonymous değişimlere, sağlıksız ve pazar değeri kayıplarına yol 2016). Sürekli yapılan bilimsel çalışmalar sayesinde açabilirler (Arslan vd. 1995; Scholz 1999). Yine bazı gerek tüketimlik ve gerekse hobi anlamında birçok nematod ve cestodların insan sağlığına zarar tür yetiştiricilik yelpazesinde yerini almaktadır. verebildiği bilinmektedir (Olson 1986).
Emre ve Kubilay 2019 - LimnoFish 5(1): 60-69 61
Özellikle Eğirdir Gölü ülkemizin tatlı su faunası Walsh ve Maloy 2008). Ayrıca, Parazit için oldukça önemli bir göldür. Eğirdir Gölü’ne enfeksiyonlarının mevsimlere ve cinsiyetine göre yapılan plantasyonlardan dolayı bugün 7 tür yerli, 6 değişimleri Quantitative Parasitology 3.0 (Rózsa vd. tür de yabancı olmak üzere 13 tür balık 2000; Reiczigel ve Rózsa 2005) programına göre bulunmaktadır (Yerli vd. 2013). 1915-2007 yılları sınanmıştır. C. pestai’de bulunan bir tür parazitin arasında Eğirdir Gölün’deki ihtiyofaunal SEM görüntüleri alınmıştır. Görüntüleme işlemleri değişimlerin değerlendirildiği bir çalışmada Eğirdir Akdeniz Üniversitesi Tıp Fakultesi Elektron bıyıklısı (Capoeta pestai)’nin kritik düzeyde Mikroskop Görüntü Analiz Ünitesi (TEMGA)’nde bulunduğu ifade edilmiştir (Küçük vd. 2009). Hem yapılmıştır. korunması ve hem de üretilmesi konusunda çalışmaların yapıldığı göl ve gölü besleyen akarsu habitatlarına uyum sağlayan Capoeta cinsine ait C. pestai türüne yönelik birçok çalışma yapılmıştır (Küçük vd. 2007; Özen vd. 2008; Ayyıldız vd. 2015; Emre vd. 2016; Demir 2017). Ancak bu türün parazitleri üzerine detaylı herhangi bir çalışma yapılmamıştır. Çalışmamızın amacı Çayköy Deresinde yaşayan bu konaktaki metazoan parazit faunasını mevsimsel olarak bazı değişkenlere göre incelemektir.
Materyal ve Metot Balık materyalinin avlandığı Çayköy Deresi, Şekil 1. Çayköy Deresi Eğirdir Gölü su toplama havzası (drenaj alanı) içindedir. Dere, Eğirdir Gölü’nün güneydoğusundaki Yılgıncak Köyü yakınlarından başlayıp, Çayköy’e kadar devam eden ve derin olmayan bir vadi içerisinde bulunur (37º 49´ 24´´K-030º 56´ 46´´D) (Şekil 1). Çayköy’den sonra Aksu HES çıkış suyu ile birleşerek yapay bir kanal ile Eğirdir Gölü’ne boşalır. Kış ve bahar aylarında 1,5-5 m3 su taşıyabilir. Yaz aylarında suları 1m3 altındadır. Çalışma için Çayköy deresinden ilk balık örnekleri ilkbahar mevsiminde alınmıştır. Balıklar elektroşokerle avlanmıştır. Araştırma süresince, toplam 150 balık bireyi incelenmiştir. Mart 2013- Şubat 2014 tarihleri arasında avlanan balıkların Şekil 2. Capoeta pestai öncelikle total boy ve ağırlıkları ölçülmüştür. Daha sonra deri, yüzgeçleri ve gözler incelenip ektoparazit Bulgular taraması yapılmıştır. Bu işlem tamamlandıktan sonra, Mart-2013/Şubat-2014 tarihleri arasında ilk önce balığın solungaçları ile iç organları mevsimsel olarak, Çayköy deresinden çıkartılmış ve stereo mikroskop altında parazitler (Eğirdir-Isparta) elektroşokerle yakalanan C. pestai aranmıştır. Bulunan parazitlerin tür, yerleşim ve türüne ait 150 bireyden 93’ü erkek, 57’si ise sayıları kaydedilip, türlere göre petri kaplarına dişi bireylerden oluşmuştur. Yapılan parazit konulmuştur. Parazitlerden bir kısmı hemen ve canlı muayenelerinde Platyhelminthes grubundan bir tür olarak incelemeye tabii tutulmuştur. Diğerleri ise digenean (Allocreadium isoporum, Loos, 1894) daha sonraki çalışmalar için % 70’lik etil alkolde ve Nemathelminthes grubundan bir tür nematod saklanmıştır. Öte yandan Nematodlar gliserin alkolde (Rhapdochona denudata, Dujardin, 1845) temizlenmiş ve % 70’lik alkolde fikse edilmişlerdir. bulunmuştur (Şekil 3, 4 ve 5). Dört mevsimdeki Parazitlerin teşhisleri Bychovskaya-Pavlovskaya muayenelerde toplam 188 A. isoporum, 1962, Gussev 1985, Gussev vd. 1987, Markevic 592 adet R. denudata paraziti saptanmıştır. 1951’e göre yapılmıştır. Parazitlerin boyama ve C. pestai türünde karşılaşılan her iki parazit türlerine tespit işlemlerinde Fernando vd. (1972)’nin ait mevsimsel yaygınlık (%), ortalama geliştirdiği yöntemden yararlanılmıştır. Konak balık yoğunluk, ortalama bolluk ve toplam parazit örneklerinin yaş tayinleri için otolitlerden sayıları Çizelge 1’de verilmiştir. Buna göre faydalanılmıştır (Murray 1994; Campana vd. 2003; A. isoporum için en yüksek yaygınlık (%52,5),
62 Emre ve Kubilay 2019 - LimnoFish 5(1): 60-69 ortalama yoğunluk (4,57) ve ortalama bolluk yaygınlık (%60) kış, ortalama yoğunluk (9,09) yaz ve (2,4) değerleri ile kış mevsiminde bulunmuştur. ortalama bolluk ise yine kış mevsimlerinde Buna karşın; R. denudata için ise, en yüksek bulunmuştur. Çizelge 1. C. pestai türündeki mevsimsel digean ve nematod gruplarına ait parazitik enfeksiyon değerleri. Enfekte Toplam İncelenen Yaygınlık Ortalama Ortalama Mevsimler Türler balık Parazit balık sayısı (%) Yoğunluk Bolluk sayısı Sayısı A.isoporum (n:42) 22 52,4 2,23 1,17 49 İlkbahar R.denudata (n:42) 24 57,1 6 3,43 144 A.isoporum (n:33) 6 18,2 2,67 0,48 16 Yaz R.denudata (n:33) 11 33,3 9,09 3,03 100 A.isoporum (n:35) 8 22,9 3 0,69 27 Sonbahar R.denudata (n:35) 18 51,4 8,11 4,17 146 A.isoporum (n:40) 21 52,5 4,57 2,4 96 Kış R.denudata (n:40) 24 60 8,42 5,05 202
(a)
Şekil 3. C. pestai’de bulunan Allocreadium isoporum’un genel görünüşü (x40).
(a)
(b)
(b) Şekil 5. C. pestai’de bulunan Nematoda (Rhapdochona denudata)’nın SEM görüntüleri (a) Genel görünüm, (b). Anterior. C. pestai türünde dört mevsim boyunca yapılan çalışmalarda A. isoporum paraziti 57 dişi balıkta değerlendirilmiştir. En yüksek yaygınlık (%) 78,9 Şekil 4. (a) R. denudata’nın genel görünüşü, (b) anteriör ortalama yoğunluk 4,93 ve 3,89 ortalama bollukla kış kısmı. mevsiminde bulunmuştur. R. denudata için ise en
Emre ve Kubilay 2019 - LimnoFish 5(1): 60-69 63 yüksek yaygınlık (%) 73,7 ile dişilerde kış ortalama bollukla ilkbahar mevsiminde yüksek mevsiminde, ortalama yoğunluk 14,50 ve 7,25 değerler bulunmuştur. Buna karşılık, R. denudata ortalama bollukla yaz mevsiminde en yüksek için ise en yüksek yaygınlık (%) 58,06 erkeklerde değerler bulunmuştur. Buna karşılık tüm ilkbaharda, ortalama yoğunluk 11,4 ile ve 5,43 mevsimlerde toplam 93 erkek balık örneğinde de ortalama bollukla kış mevsiminde yüksek değerler değerlendirme yapılmıştır. Buna göre; A. isoporum bulunmuştur. A. isoporum ve R. denudata’ya ait için en yüksek yaygınlık (%) 48,40 erkeklerde mevsim ile konak cinsiyetlerine göre dağılımlarına ilkbaharda, ortalama yoğunluk 3,67 ile kış ve 1,13 ait veriler Çizelge 2’de verilmiştir.
Çizelge 2. C. pestai türünün cinsiyetlere göre mevsimsel digenean ve nematod gruplarına ait parazitik enfeksiyon değerleri.
İncelenen Parazitli Toplam Yaygınlık Ortalama Ortalama Mevsimler Cinsiyet Türler balık balık Parazit (%) Yoğunluk Bolluk sayısı sayısı Sayısı
A.isoporum (n:11) 7 63,6 2 1,27 14 Dişi R.denudata (n:11) 6 54,5 2,83 1,55 17 İlkbahar A.isoporum (n:31) 15 48,4 2,33 1,13 35 Erkek R.denudata (n:31) 8 58,06 7,06 4,1 127
A.isoporum (n:8) 2 25 4 1 8 Dişi R.denudata (n:8) 4 50 14,5 7,25 58 Yaz A.isoporum (n:25) 4 16 2 0,32 8 Erkek R.denudata (n:25) 7 28 6 1,68 42
A.isoporum (n:19) 4 21,1 2,75 0,58 11 Dişi R.denudata (n:19) 9 47,4 8,78 4,16 79 Sonbahar A.isoporum (n:16) 5 31,3 3,2 1 16 Erkek R.denudata (n:16) 9 56,3 7,44 4,19 67
A.isoporum (n:19) 15 78,9 4,93 3,89 74 Dişi R.denudata (n:19) 14 73,7 6,29 4,63 88 Kış A.isoporum (n:21) 6 28,6 3,67 1,05 22 Erkek R.denudata (n:21) 10 47,6 11,4 5,43 114
A. isoporum’un mevsimlere göre konak C. pestai için mevsimsel olarak yapılan yaş grupları değerlendirildiğinde, ilkbahar örneklemelerde nematod grubundan R. denudata döneminde III. Yaş grubuna ait balıkların A. parazitine tüm mevsimlerde rastlanmıştır (Çizelge isoporum ile enfazla enfekte olduğu görülmüşütür. 1,2 ve 5,6). Çizelge 5’de yaş grubuna göre yapılan (Çizelge 3). Diğer değerlendirme sonuçları aynı değerlendirmede en fazla enfekte balık sayısının kış Çizelge’de verilmiştir. Yine boya göre değerlendirme mevsiminde III yaşındaki bireylerde olduğu de Çizelge 4’de gösterilmiştir. Buna göre görülmüştür. Yine Çizelge 6’da boya göre yapılan ilkbahardaki 14,1-18,0 cm boy grubunda daha fazla değerlendirmede ise 14,1-18,0 grubundaki konak örneğin A. isoporum ile enfekte olduğu görülmüştür. örnekleinin daha fazla enfekte olduğu görülmüştür. Ayrıca mevsimsel olarak değerlendirilen Çizelge 5 ve 6’da Capoeta pestai’de bulunan örneklemelerin boya göre değerlendirmeleri Çizelge R. denudata’nın konak boy ve yaş gruplarına göre 4’de verilmiştir. dağılımları ile ilgili diğer sonuçlar verilmiştir.
64 Emre ve Kubilay 2019 - LimnoFish 5(1): 60-69
Çizelge 3. C. pestai’de bulunan Allocreadium isoporum’un konak yaş gruplarına göre değerlendirilmesi. Yaş Grupları O I II III IV V VI VII VIII IX X
İlkbahar
İncelenen Balık Sayısı 2 2 6 15 10 3 1 0 3
Enfekte Balık Sayısı 0 0 2 10 6 2 1 0 1
Yaygınlık (%) 0 0 33,3 66,7 60 66,7 100 0 33,3
Ortalama Yoğunluk 0 0 1,5 2 2 4,5 3 0 2
Ortalama Bolluk 0 0 0,5 1,33 1,2 3 3 0 0,67
Toplam Parazit Sayısı 0 0 3 20 12 9 3 0 2
Yaz
İncelenen Balık Sayısı 8 12 3 4 3 2 1 0 0
Enfekte Balık Sayısı 1 3 0 0 1 1 0 0 0
Yaygınlık (%) 12,5 25 0 0 33,3 50 0 0 0
Ortalama Yoğunluk 2 3 0 0 1 4 0 0 0
Ortalama Bolluk 0,25 0,75 0 0 0,33 2 0 0 0
Toplam Parazit Sayısı 2 9 0 0 1 4 0 0 0
Sonbahar
İncelenen Balık Sayısı 2 4 4 3 4 6 3 0 8 0 1
Enfekte Balık Sayısı 1 0 1 0 1 1 2 0 3 0 0
Yaygınlık (%) 50 0 25 0 25 16,7 66,7 0 37,5 0 0
Ortalama Yoğunluk 3 0 1 0 3 3 2 0 4,33 0 0
Ortalama Bolluk 1,5 0 0,25 0 0,75 0,5 1,33 0 1,63 0 0
Toplam Parazit Sayısı 3 0 1 0 3 3 4 0 13 0 0
Kış
İncelenen Balık Sayısı 5 8 6 11 3 3 0 0 8 0 0
Enfekte Balık Sayısı 0 1 2 9 2 3 0 0 4 0 0
Yaygınlık (%) 0 12,5 33,3 81,8 66,7 100 0 0 50 0 0
Ortalama Yoğunluk 0 2 6,5 3,67 6 3 0 0 6,75 0 0
Ortalama Bolluk 0 0,25 2,17 3 4 3 0 0 3,38 0 0
Toplam Parazit Sayısı 0 2 13 33 12 9 0 0 27 0 0
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Çizelge 4. C. pestai’de bulunan Allocreadium isoporum’un konak boy gruplarına göre değerlendirilmesi.
I II III IV V Boy Grupları ( cm) (6,0-10,0 cm) (10,1-14,0 cm) (14,1-18,0 cm) (18,1-22,0 cm) (22,1-28,0 cm)
İlkbahar
İncelenen Balık Sayısı 2 8 17 15 0
Enfekte Balık Sayısı 0 2 12 8 0
Yaygınlık (%) 0 25 70,6 53,3 0
Ortalama Yoğunluk 0 1,5 2,08 2,63 0
Ortalama Bolluk 0 0,38 1,47 1,4 0
Toplam Parazit Sayısı 0 3 25 21 0
Yaz
İncelenen Balık Sayısı 7 14 7 5 0
Enfekte Balık Sayısı 1 3 1 1 0
Yaygınlık (%) 14,3 21,4 14,3 20 0
Ortalama Yoğunluk 2 3 1 4 0
Ortalama Bolluk 0,29 0,64 0,14 0,8 0
Toplam Parazit Sayısı 2 9 1 4 0
Sonbahar
İncelenen Balık Sayısı 0 9 6 12 7
Enfekte Balık Sayısı 0 1 1 4 2
Yaygınlık (%) 0 11,1 16,7 33,3 28,6
Ortalama Yoğunluk 0 1 3 3,5 3
Ortalama Bolluk 0 0,11 0,5 1,17 0,86
Toplam Parazit Sayısı 0 1 3 14 6
Kış
İncelenen Balık Sayısı 6 10 15 5 4
Enfekte Balık Sayısı 0 1 11 5 4
Yaygınlık (%) 0 10 73,3 100 100
Ortalama Yoğunluk 0 2 4,18 4,6 6,25
Ortalama Bolluk 0 0,2 3,07 4,6 6,25
Toplam Parazit Sayısı 0 2 46 23 25
66 Emre ve Kubilay 2019 - LimnoFish 5(1): 60-69
Çizelge 5. Capoeta pestai’de bulunan Rhapdochona denudata’nin yaş gruplarına göre değerlendirilmesi.
Yaş Grupları O I II III IV V VI VII VIII IX X
İlkbahar
İncelenen Balık Sayısı 2 2 6 15 10 3 1 0 3 0 0
Enfekte Balık Sayısı 1 2 3 11 4 2 1 0 0 0 0
Yaygınlık (%) 50 100 50 73,3 40 66,7 100 0 0 0 0
Ortalama Yoğunluk 1 3,5 1,33 9,82 1,5 8 2 0 0 0 0
Ortalama Bolluk 0,5 3,5 0,67 7,2 0,6 5,33 2 0 0 0 0
Toplam Parazit Sayısı 1 7 4 108 6 16 2 0 0 0 0
Yaz
İncelenen Balık Sayısı 8 12 3 4 3 2 0 0 0 0 0
Enfekte Balık Sayısı 2 4 2 1 2 0 0 0 0 0 0
Yaygınlık (%) 25 33,3 66,7 25 66,7 0 0 0 0 0 0
Ortalama Yoğunluk 6,5 6,25 12 25 6,5 0 0 0 0 0 0
Ortalama Bolluk 1,63 2,08 8 6,25 4,33 0 0 0 0 0 0
Toplam Parazit Sayısı 13 25 24 25 13 0 0 0 0 0 0
Sonbahar
İncelenen Balık Sayısı 2 4 4 3 4 6 3 0 8 0 1
Enfekte Balık Sayısı 0 2 3 0 2 5 3 0 3 0 0
Yaygınlık (%) 0 50 75 0 50 83,3 100 0 37,5 0 0
Ortalama Yoğunluk 0 7 4,67 0 7,5 8,4 5 0 15,33 0 0
Ortalama Bolluk 0 3,5 3,5 0 3,75 7 5 0 5,75 0 0
Toplam Parazit Sayısı 0 14 14 0 15 42 15 0 46 0 0
Kış
İncelenen Balık Sayısı 5 8 6 11 3 3 0 0 8 0 0
Enfekte Balık Sayısı 2 1 2 11 2 2 0 0 4 0 0
Yaygınlık (%) 40 12,5 33,3 100 66,7 66,7 0 0 50 0 0
Ortalama Yoğunluk 35,5 2 5,5 7,09 8 3,5 0 0 4,25 0 0
Ortalama Bolluk 14,2 0,25 1,83 7,09 5,33 2,33 0 0 2,13 0 0
Toplam Parazit Sayısı 71 2 11 78 16 7 0 0 17 0 0
Emre ve Kubilay 2019 - LimnoFish 5(1): 60-69 67
Çizelge 6. Capoeta pestai’de bulunan Rhapdochona denudata’nin boy gruplarına göre değerlendirilmesi.
I II III IV V Boy Grupları (cm) (6,0-10,0 cm) (10,1-14,0 cm) (14,1-18,0 cm) (18,1-22,0 cm) (22,1-28,0 cm)
İlkbahar
İncelenen Balık Sayısı 2 8 17 15 0
Enfekte Balık Sayısı 1 5 12 6 0
Yaygınlık (%) 50 62,5 70,6 40 0
Ortalama Yoğunluk 1 2,2 9,17 3,67 0
Ortalama Bolluk 0,5 1,38 6,47 1,47 0
Toplam Parazit Sayısı 1 11 110 22 0
Yaz
İncelenen Balık Sayısı 7 14 7 5 0
Enfekte Balık Sayısı 2 5 3 1 0
Yaygınlık (%) 28,6 35,7 42,9 20 0
Ortalama Yoğunluk 6,5 8 12,33 10 0
Ortalama Bolluk 1,86 2,86 5,29 2 0
Toplam Parazit Sayısı 13 40 37 10 0
Sonbahar
İncelenen Balık Sayısı 0 9 6 12 7
Enfekte Balık Sayısı 0 5 2 8 3
Yaygınlık (%) 0 55,6 33,3 66,7 42,9
Ortalama Yoğunluk 0 5,6 7,5 7,13 15,33
Ortalama Bolluk 0 3,11 2,5 4,75 6,57
Toplam Parazit Sayısı 0 28 15 57 46
Kış
İncelenen Balık Sayısı 6 10 15 5 4
Enfekte Balık Sayısı 2 1 13 5 3
Yaygınlık (%) 33,3 10 86,7 100 75
Ortalama Yoğunluk 35,5 2 6,85 5 5
Ortalama Bolluk 11,8 0,2 5,93 5 3,75
Toplam Parazit Sayısı 71 2 89 25 15
68 Emre ve Kubilay 2019 - LimnoFish 5(1): 60-69
Tartışma ve Sonuç ilkbahar (% 23) ve yaz mevsimlerinde (% 16,7) Ülkemizdeki endemik balıklardan C. pestai yaygınlık oranı ile karşılaşılmıştır. Sonuç olarak türünde bulunan helmint biyoçeşitliliğinin yukarıda belirtildiği gibi; Çayköy Deresindeki C. incelendiği bu çalışmamızda, C. pestai türünün % pestai’de bulunan iki parazit türü konak için yeni 58,0’nın en az iki tür parazitle enfekte olduğu kayıt özelliğini taşımaktadır. Bu nedenle, gerek saptanmıştır. Zayıf bir patojen olarak bilinen ve mevcut alandaki ve gerekse Eğirdir Gölü’ndeki özellikle sazangiller ailesine mensup türlerde stoklara ait tehditlerin tespiti ve yine Türkiye’nin barsakta bulunan A. isoporum türü çalışmamızda bu helmint biyoçeşitliliğinin belirlenmesi noktasında konakta bulunmuştur. Bu konak için yeni kayıttır. C. çalışmamız önem arz etmektedir. pestai türünde ağırlıklı olarak yakalanan balıkların 1/3’ü dişi bireylerden oluşmuştur. En yüksek Teşekkür yaygınlık oranları ilkbahar ve kış örneklemelerinde Bu makale Süleyman Demirel Üniversitesi, Fen bulunmuştur (% 52,5). En düşük ise yaz Bilimleri Enstitüsünde “Akdeniz Bölgesi’ndeki örneklemelerinde belirlenmiştir (% 18,2). Buna Capoeta erhani, Capoeta pestai, Capoeta mauricii karşılık, III yaşındaki ilkbahar örneklerinde en fazla Türlerinin Helmint Parazitlerinin Biyoçeşitliliğinin enfekte balığa rastlanmıştır. Moravec (1992) Tuna Araştırılması” başlıklı tezden üretilmiştir. havzasındaki Rokytna Nehirinde Leuciscus cephalus konağına yönelik yaptığı çalışmada en yüksek Kaynaklar yaygınlık oranını ilkbahar aylarında (%100), ay Anounymous 2016. The state of world fisheries and ortalama yaygınlık oranını ise % 73 olarak aquaculture. FAO Fisheries and Aquaculture saptamıştır. Koyun (2001), Enne Baraj Gölünde Department. p190. yaptığı çalışmada Alburnus alburnus konağında Arslan A, Dinçoğlu A, Güven A. 1995. Su ürünleri ile insanlara geçen hastalıklar. Kafkas Üniv Vet Fak Mayıs ve Haziran (1998)’da %15 ve %19; Nisan ve Derg. 1(1-2):103-107. Mayıs (1999)’da ise %15 ile %10 yaygınlık oranında Aydogdu A, Altunel FN, Yıldırımhan HS. 2002. The A. isoporum paraziti belirlemiştir. Aydoğdu vd. Occurrence of Helminths Parasites in Barbel (Barbus (2002), Doğancı Baraj Gölündeki Barbus plebejus Plebejus Escherichi Steındacher, 1897) of the konağının helminth faunasına yönelik yaptıkları Dogancı (Bursa) Dam Lake, Turkey. Acta Vet- araştırmada A. isoporum parazitine Mayıs ve Haziran Beograd, 52:(5-6):369380, UDK 619 616 995 1. aylarında %75 ve %33,3; Eylul ve Ekim aylarında Aydoğdu A, Emre Y, Emre N, Altunel FN. 2011. The %11,1 ve %20 ve toplamda ise %19,1 yaygınlık Occurrence of Helminth Parasites (Nemathelminthes) oranında rastlanılmıştır. Yine Raissy ve Ansari in Some Freshwater Fish from Streams Discharging (2012) İran’da Arman Nehiri balıklarındaki into Antalya Bay in Antalya, Turkey: Two New Host Records from Antalya. Turk J Zool,35: (6). 859-864. parazitlere yönelik yaptıkları çalışmada C. aculeata, doi:10.3906/zoo-0912-16 C. damascina ve Barbus barbulus’un barsaklarında Ayyıldız H, Emre Y, Yağcı A, Altin A. 2015. Length– yaygınlık oranları %2, %0,79 ve % 1,4 şeklinde Weight Relationships of Eight Freshwater bulmuşlardır. (Cypriniformes) Fish Species from Turkey. J Appl Diğer yandan C. pestai konağında, nematod Ichthyol. 1: 1,1-2. grubuna ait bir tür olan Rhabdochona denudata türü doi: 10.1111/jai.12828 bulunmuştur. Bu tür de yeni kayıt özelliği Bychovskaya - Pavlovskaya IE. 1962. Key to Parasites of taşımaktadır. Bu türle alakalı olarak ayrıca SEM Freshwater Fishes of the U.S.S.R. Moskova – çalışması yapılmıştır. C. pestai’de en yüksek Leningrad: Izdatel’stvo Akademi Nauk SSR. (In yaygınlık oranı % 58,1 ile ilkbahar, en düşük ise %28 Russian: English Translation – Israel Program for Scientific Translation), Jerusalem, p.919. ile yaz örneklemelerindeki erkek bireylerde Campana SE, Stanley RD, Wischniowski S. 2003. saptanmıştır. III. yaş gruplarında (kış) ve 14,1-18,0 Suitability of Glycerin-Preserved Otoliths for Age boy aralığındaki bireylerde daha fazla enfekte olan Validation Using Bomb Radiocarbon. J Fish Biol, 63, bireylere rastlanmıştır. Aydoğdu vd., (2011) Antalya 848-854. Körfezine dökülen akarsularda yaptıkları çalışmada doi: 110.1046/j.1095-8649.2003.00174.x sınırlı sayıdaki Capoeta antalyensis’te %86,6 Demir O. 2017. Endemik İçsu Balıkları Yetiştiriciliğinin yaygınlık oranında R. denudata tespit edilmiştir. Önemi: Eğirdir Sirazı (Capoeta pestai Pietschmann, Pazooki vd. (2012) İran-Kerman ilindeki dört ayrı su 1933), Süleyman Demirel Üniversitesi Eğirdir Su kaynağında avlanan C. damascina’da yaygınlık Ürünleri Fakültesi Dergisi, 13(1), 88-98. E-Dergi oranını %73,39; Cyprinion watsoni’da %88,60, ISSN: 1308 – 7517. Emre Y, Altın A, Ayyıldız H, Dölcü B, Küçük F, Özen Ö. Schistura sargadensis’da %2.94 ve Channa 2016. Age and Growth of Capoeta pestaı gachua’da ise % 13,63 şeklinde R. denudata (Actınopterygıı: Cypriniformes: Cyprinidae) in A bulmuşlardır. Koyun vd. (2015) Murat Nehirin’de Small River Enterıng Lake Eğirdır, Turkey, Acta yaşayan B. lecerta konağında bu nematod da sadece Ichthyologica Et Piscatoria, 46: (2) 57–63.
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doi: 10.3750/AIP2016.46.2.01 Seafood Quality Determination, Proceedings of an Fernando CH, Furtado JI, Gussev AV, Hanek G - Kakonge İnternational Symposium Coordinated by the S. A. 1972. Methods for the Study of Freshwater Fish University of Alaska Sea Grant Collage Program, Parasites. Deparement of Biology, University of Anchorage, Alaska, USA, 10-14. Waterloo, Waterloo, Ontario, Canada. p.76. Özen MR, Turna İ, Cınar K. 2008. As an Example of Gussev AV. 1985. In: Key to Parasites of the Freshwater Ecological Devastation Egırdır Lake (Turkey), BJAS, Fishes of the USSR. Fauna, vol. 2. (ed. By ON Bauer) 14: (2), 195-200. Publ. House Nauka. Leningrad, 1 - 418 p. Pazooki J, Masoumian M. 2012. Synopsis of the Parasites Gussev AV, Poddubnaya AV, Abdeeva VV. 1987. Key to in Iranian Freshwater Fishes, IRAN J FISH SCI, 11: Parasites of the Freshwater Fishes of the USSR. (3), 570-589. Fauna, vol. 3 (ed. By ON Bauer) Publ. House Nauka. Raissy M, Ansari M. 2012. Parasites of Some Freshwater Leningrad, 1 - 532 p. Fish from Armand River, Chaharmahal va Bakhtyari Koyun M. 2001 Enne Baraj Gölündeki Bazı Balıkların Province, Iran. Iranian J Parasitol, 7: (1), 73-79. Helmint Faunası (Doktora Tezi, Uludağ Üniversitesi, Reiczigel, J, Zacarias I, Rósza, L., (2005), A Bootstrap 119 sayfa. Test of Stochastic Equality of Two Populations. The Koyun M, Ulupınar M, Gül A. 2015. Seasonal Distribution American Statistician, 59, 156–161. of Metazoan parasites on Kura Barbell (Barbus Rózsa, L., Reiczigel, J., Majoros, G. (2000). Quantifying lacerta) İn Eastern Anotolia, Turkey, Pakistan J Zool, Parasites in Samples of Hosts. J Parasitol, 86: 228– 47: (5), 1253-1261. 232. Kuru M, Yerli S, Mangit F, Ünlü E, Alp A. 2014. Fish Scholz T. 1999. Parasites in Cultured and Feral Fish. Biodiversty in İnland Waters of Turkey, Journal of Veterinary Parasitology 84, 317–335. Academic Documents for Fisheries and Aquaculture, doi: 10.1016/S0304-4017 (99) 00039-4 3: 93-120. Walsh MG, Maloy AP. 2008. Comparison of Rainbow Küçük F, Turna İ, Demir O. 2007. Taxonomic Smelt Age Estimates from Fin Rays and Otoliths. N Characteristics and Distribution of Capoeta pestai Am J Fish Manag 28, 42-49. (Pietschmann, 1933) (Pisces: Cyprinidae). Süleyman doi: 10.1577/MO6-292.1 Demirel University Journal of Natural and Applied Yerli SV, Alp A, Yeğen V, Uysal R, Yağcı MA, Balık İ. Sciences, 11: 18-25. 2013. Evaluation of the Ecological and Economical Küçük F, Sarı HM, Demir O, Gülle İ, Güçlü SS, Gümüş E. Results of the Introduced Alien Fish Species in Lake 2009. Review of The İchthyofaunal Changes İn Lake Eğirdir, Turkey, TURK J FISH AQUAT SC, 13: 795- Eğirdir Between 1915 And 2007. TURK J ZOOL, 33: 809. 277-286. doi :10.4194/1303-2712-v13_5_03 doi:10.3906/zoo-0811-16 Yıldırım Ö, Mazlum MD, Güllü K. 2002. Doğu Karadeniz Markevic AP. 1951. Parasitic Fauna of Freshwater Fish of bölgesinde kullanılan bazı ticari yemlerin gökkuşağı the Ukrainian SSR.İsrael Program Scientific alabalığının (Oncorhynchus mykiss W.,1792) Translations, Jarusalem, 10–11. biyoekonomisi üzerine etkisi. YYÜ Ziraat Fakültesi Moravec F. 1992. Observation on The Bionomy of Tarım Bilimleri Dergisi (J. Agric. Sci.), 12(1): 7-12. Allocreadium İsoporum (Loos,1894). (Trematoda: Yılmaz E. 1998. Gökkuşağı alabalığı yetiştiriciliğinde Allocreadiidae), FOLIA PARASIT, 39: 133-144. farklı oranlarda ekstrüde yem kullanımının balıkların Murray CB. 1994. A Method For Preparing Chinook gelişimine etkileri [Yüksek Lisans Tezi]. Ondokuz Salmon Otoliths For Age-Determination, and Mayıs Üniversitesi. 46s. Evidence of Its Validity. Trans Am Fish Soc, 123, Yiğit M, Aral O. 1999. Gökkuşağı alabalığının 358-367. (Oncorynchus mykiss Walbaum, 1792) tatlısu ve deniz Olson RE. 1986. Marine Fish Parasite of Public Health suyundaki büyüme farklılıklarının karşılaştırılması. Importance. D.E. Kramer and J. Liston (Editors), Turk J Vet Anim Sci, 23: 53-59.
LIMNOFISH-Journal of Limnology and Freshwater Fisheries Research 5(1): 70-75 (2019)
Assessment of Fish Exports from Blantyre District, Southern Malawi
Langson SAMALA* Fanuel KAPUTE
Department of Fisheries & Aquatic Sciences, Mzuzu University, Private Bag 201, Mzuzu 2, Malawi
ABSTRACT ARTICLE INFO
This study presents findings on fish exports from Blantyre, which is the biggest SHORT COMMUNICATION commercial city in Malawi. A cross-sectional research design was used in the study in which fish quantities, species, processing methods, gender of fish Received : 20.02.2018 exporters and export destinations were analysed. Non-probability data collection Revised : 08.08.2018 methods were used on the secondary data that was collected from Blantyre Accepted : 06.09.2018 District Fisheries Office. Findings indicate that a total of 9596 kg (9.6 metric tonnes) of fish was exported within a period of six months in the year 2013 Published : 25.04.2019 comprising all species available in the local markets and those endemic in Malawi’s water bodies. Most fish were exported as sun-dried, para-boiled and DOI:10.17216/LimnoFish.397020 smoked. Findings suggest that fish exports from Malawi are in their infancy, fluctuate, and are insignificant for making a positive and sustainable impact to the * CORRESPONDING AUTHOR economy of the country. It is recommended that national and regional policies [email protected] should be fully utilized to curb exports of fish in order to sustainably satisfy the Phone : +265 995 388 420 local huge demand for fish. About 82.6% of the fish exporters were women underpinning the need for policy considerations that value their critical role in the fish export trade such as women empowerment. Keywords: Fish exports, species, processing, destinations, gender
How to Cite Samala L, Kapute F. 2019. Assessment of Fish Exports from Blantyre District, Southern Malawi. LimnoFish. 5(1): 70-75. doi: 10.17216/LimnoFish.397020
Introduction exported fish to neighbouring Tanzania, In Malawi, documented fish exports are Mozambique and Zambia between 2015 and 2016 estimated at 0.102 metric tonnes, accounting for a with an estimated annual quantity of 24115.68 metric monetary value of MWK (Malawi Kwacha currency) tonnes valued at 41.6 million dollars (Mussa et al. 22,000 (US$ 250) in 1998 to 159.5 metric tonnes 2017). Such reports exist at a time when the country valued at MWK18.5 million (US$170,000) in 2002 was in a dire shortage of fish supply with an average (Commonwealth/GTZ 2007). The figures indicate per capita annual fish consumption estimated at 5.6 that fish exports increased in the period of 1998 to kg, far below the 13-15 kg recommended by the 2002. However, the Government of Malawi (GoM World Health Organisation (Kapute 2017). The 2012) reported that annually, an average of 86 tonnes demand for fish in Malawi has in recent years of fish was exported between 2002 and 2010. Reports tremendously increased due to increased population also indicated that quantities of annual fish exports against the declining fish catches from the natural within the same period greatly fluctuated due to waters. In Malawi, fish provides the best and most dwindling fish harvests (Commonwealth/GTZ 2007; affordable source of dietary animal protein to many Donda and Njaya 2007; GoM 2011, 2012). Fish people contributing about 28% of all dietary animal catches from Malawi were estimated at 70000 and protein supply (Kapute 2017). Another concern is 3000 metric tonnes per annum from capture fisheries that there is little consideration about quality and fish farming, respectively (GoM 2012). To the standards for fish trade in Malawi and clearly contrary, the domestic demand for fish in the country indicating that fish quality standards have not been was estimated at around 110000 metric tonnes per well developed for exports to highly regulated year (GoM 2012) indicative of a supply deficit. markets (Kapute 2009; GoM 2012; Manyungwa- Recent reports showed that Malawi informally Pasani et al. 2017). The traders have thus, been
Samala and Kapute 2019 - LimnoFish 5(1): 70-75 71 trading without proper licensing and no 2012). Data collected for this study did not indicate empowerment. In addition, there have been limited the exact quantities exported per species and as such, studies focusing on the exports of fish from Malawi. frequencies were used. The monthly fish This study differs from earlier studies because it exports were added for all months from May to focuses on formal fish exports. The novelty of this October 2013, and quantities were also added to give study provides a foundation to the sustainability of a total quantity of fish exported within the period the fish exports in order for the country to benefit of study. The number of fish exporters were from the trade and diversify the income generating separately classified into three categories according activities in the fisheries value chain. In this study, to the quantity of fish exported and percentages exports of fish from Malawi were assessed in order of monthly quantities from total quantity exported to assist the government in establishing the were computed. The classes were: less than 50 kg magnitude of fish quantities exported, fish processing (fq<50kg), between 50 and 100 kg (50 < fq < 100), methods, fish species, gender of exporters and fish and greater than 100 kg (fq>100), where fq is export destinations. the fish quantity exported. The percentage of traders in each category was calculated. To determine Material and Methods fish processing methods used for fish destined Study area and data collection for trading in the international markets, the Cross-sectional research design was used in the proportion of each processed method of the fish study (Bhattacherjee 2012) and fish export data were species was summarized in the bar graphs to observe collected from Blantyre District Fisheries Office in the frequently exported forms of processed fish. southern Malawi. Blantyre is the commercial and Gender of fish exporters and destinations of industrial city of Malawi and forms the fish exports communication and transportation hub to all parts of The percentages of male and female fish the country as well as the neighbouring countries. exporters were found for each month of the study to Fish exports data are collected by the Malawi understand whether they were equal percentages for Department of Fisheries when issuing the each month. All countries, which serve as markets for phytosanitary certificates to fish exporters. fish from Blantyre, were identified and frequencies Information that is contained in the phyto-sanitary of fish traders were calculated and recorded. This certificates includes destinations, fisheries quality method was preferred due to its robustness control unit, name and address of exporter, declared in encompassing specific details without overlooking name and address of consignee, declared quality and the importance of the details. name of fish product, scientific name of fish, number and description of packages, declared means of Statistical analysis conveyance and treatment, among others. The Data were analyzed in SPSS Software version Department of Fisheries started providing the phyto- 16.0. Chi – square (goodness of fit) test was used to sanitary certificates to Blantyre District Fisheries test for the differences in the distributions Office in March 2013. To ensure validity and of categories of fish exporters and their gender. The reliability of the data collected, 10 duplicate sanitary hypotheses were tested at 5% significance level. certificates were collected from Blantyre District Graphs were generated using Microsoft Excel 2010. Fisheries Office before the actual data collection exercise. The collected duplicate Phyto-sanitary Results certificates were checked by qualified technical Quantity of fish exports personnel in the Department of Fisheries and A total of 9596 kg of fish were exported in the Aquatic Sciences at Mzuzu University, Malawi. Fish period studied and the highest quantity was exports data for a period of six months, from May exported in the month of June (39.2%). It should be to October, 2013 were collected from a total of 217 noted that although the percentages of fish exported individual traders using non - probability per month were fluctuating, the trend showed an sampling technique i.e. all the data available were increase from 7.9%, 12.0% and 17.5% from July, collected for the study (Ukaonu et al. 2011; August and September respectively (Figure 1). Haambiya et al. 2013). There were significant differences among the Species and quantity of fish exported three categories of fish exporters in all the months Fish species that an individual person could export studied (Table 1). The months showing the degrees were summarized as percentages instead of listing of freedom (df) of one suggest no fish exports of used in earlier studies (Jayalal and Ramachandran more than 100 kg.
72 Samala and Kapute 2019 - LimnoFish 5(1): 70-75
45,0 39,2 40,0 35,0 30,0 25,0 20,0 17,5 14,1 15,0 12,0 9,2 10,0 7,9 5,0 0,0 Percentagequantityof exported May June July August September October Month
Figure 1. Percentage of fish exports from May to October 2013 from Blantyre Table 1. Chi-square (goodness of fit) test results for the three categorised fish exporters May June July August September October 훘ퟐ − 퐯퐚퐥퐮퐞 13.300a 28.374a 4.263a 9.143a 11.789a 12.250a df 2 1 1 1 2 2 Asymp. Sig. 0.001 0.000 0.039 0.002 0.003 0.002 Values with similar superscripts in a row are significantly different (P < 0.05). Fish species on international trade (Makakana), Enteromius paludinosus Fish species that were exported are (Peters, 1852) (Matemba), and Rhamphochromis Engraulicypris sardella (Günther, 1868) (local name: species (Mcheni). However, frequently exported Usipa), Clarias gariepinus (Burchell, 1822) species were E. sardella and C. gariepinus with more (Catfish: Mlamba), Oreochromis species (Weyl et al than 75% and 65% of the fish exporters respectively 2010) (Chambo), Oreochromis mossambicus (Figure 2).
Usipa(%) Mlamba(%) Chambo(%) Makakana(%) Utaka(%) Mcheni(%) Matemba(%) Other spp(%) 100,00 80,00 60,00 40,00 20,00 0,00 May June July August September October
Percentage fish of exported Month
Figure 2. Percentages of fish exporters from Blantyre to various export destinations from May to October, 2013.
Fish processing methods while Rhamphochromis species (Mcheni) were Observed fish processing methods were sun exported in the smoked form compared to other - dried and para - boiling for E. sardella (Figure 3). forms (Figure 4). Commonly, Oreochromis species (local name: Fishes such as Barbus species (Matemba), Chambo), Diplotaxodon spp. (Ndunduma) and Copadichromis species (Utaka), Bagrus species C. garipinus (Catfish) were exported (Kampango), Buccochromis species (Mbaba) were in smoked form. A small quantity of Oreochromis mainly exported in smoked and sun - dried forms species were also exported frozen (Figure 5).
Samala and Kapute 2019 - LimnoFish 5(1): 70-75 73
Sundried Usipa Paraboiled Usipa 600 500 400 300 200
100 Quantity of fish(kg) 0 May June July August Sept Oct Nov Dec Month
Figure 3. Processed form of exported E. sardella
Fresh/frozen Chambo Smoked Chambo Smoked Mlamba Smoked Ndunduma Frozen/fresh Mcheni Smoked Mcheni 200
150
100
50
Quantity of fish(kg) 0 May June July August Sept Oct Nov Dec Month
Figure 4. Processed form of Oreochromis spp. (Chambo), Diplotaxodon spp. (Ndunduma), Rhamphochromis spp. (Mcheni) and Clarias spp. (Catfish) exported from Blantyre.
Smoked Kampango Sundried Utaka Smoked Utaka Sundried Matemba Smoked Mbaba 30 25 25 18 20 16 15 12 10 8 8 10 10 6 3 5 5 2 1
0 Quantity of fish(kg)
Month
Figure 5. Processed form of Copadichromis spp. (Utaka), Enteriomius spp.(Matemba), Bagrus meridionalis (Kampango) and Buccochromis spp. (Mbaba) exported in 2013. Export destinations of fish from Blantyre Some fish were also exported to Botswana (n=1), Results show that the Republic of South Africa Mozambique (n=1) and Vietnam (n=1) in the same (n=214) is the main importer of fish from Malaŵi. period.
74 Samala and Kapute 2019 - LimnoFish 5(1): 70-75
Contrary to the expectations, there was no export Zambia and Zimbabwe in the period under study of fish to the neighboring countries like Tanzania, (Table 2). Table 2. Number of fish exporters from Blantyre in months of study
Month Republic of South Africa (n=214) Botswana (n=1) Other (n=2)
May 20 0 0 June 98 0 1 July 19 0 0 August 27 1 0 September 19 0 0 October 31 0 1
Gender of fish exporters (P = 0.000), July (P = 0.000), August (P=0.001) About 13.8% of fish exporters were males and October (P=0.000) and the month of and 86.2% were females with a significant September showed gender difference difference (P<0.05) between the gender of was insignificant (P = 0.108) as reported fish exporters in the months of May (P = 0.000, June in Table 3. Table 2. Percentages and Chi - square (goodness of fit) test results for fish exporters.
Month Male (%) Female (%) 2-value Asymp. Sig.
May 5.0 95.0 16.200a 0.000 June 12.1 87.9 56.818a 0.000 July 5.3 94.7 15.211a 0.000 August 17.9 82.2 11.571a 0.001 September 31.6 68.4 2.579b 0.108 October 15.6 84.4 15.125a 0.000
Discussion (14.1 %) and October (17.5 %) may be indicative of The study of fish exports from Blantyre District increased fish availability on the markets following in Malawi was limited by the availability of sufficient the opening of the lakes following seasonal closure in data. Data that was available at the time of the study November and/or December to March. Highest fish were for six months only. The findings in this study exports (39.2 %) in June may be due to traders were consistent with earlier studies that fish turning up as production of E. sardella increased quantities exported from Malawi have been low along with other species. This study revealed that (Commonwealth/GTZ 2007; Donda and Njaya 2007; E. sardella and Copadichromis species which were Kapute 2009; GoM 2011, 2012). Quantities as low as commercially - less valuable fishes (GoM 2012; 9596 kg exported in six months from Blantyre Singini 2013) were the mostly exported by traders (at District may imply that fish exports at the current rate least 75% of the traders exported the two species). cannot meaningfully contribute to the economy of the The fish processing methods such as sun-drying, country. However, due to increased informal cross smoking and para-boiling indicate that the exported border trade as evidenced by the informal fish export fishes were destined for local markets (Kapute 2009; of 24115.68 metric tonnes between 2015 and 2016 GoM 2012). The processing methods also suggest from Malawi to neighboring countries (Mussa et al. that the species of fish exported are small in size. 2017), fish export quantities could be higher than It was observed fish were mainly exported to the reported. The inconsistences earlier reported by Republic of South Africa as earlier reported by GoM Kapute (2017) may rise due to poor monitoring of the (2011). Republic of South Africa imported largest fish exports by the Government of Malawi. The quantities of fish from Malawi probably due to many increasing trend of fish export quantities from May Malawians staying in that country as consumer (9.2 %), July (7.9 %), August (12.0 %), September studies in Johannesburg found that demand for fish
Samala and Kapute 2019 - LimnoFish 5(1): 70-75 75 species depends on culturally acquired tastes (Kapute policy progress; [cited 2018 Jan 04]. Available from 2017). However, other studies have established http://pubs.iclarm.net/wfcms/file/bmz/Malawi_policy reported fish exports to Zambia, Tanzania and .pdf. Zimbabwe (Kapute 2017; Mussa et al. 2017) which Government of Malawi (GoM). 2011. Annual economic were not found in the current study. The large report 2012. Lilongwe: Ministry of Development Planning and Cooperation. percentage of female fish exporters (86.2%) and the Government of Malawi (GoM). 2012. National fisheries monthly gender differences agree with the findings policy 2012-2017. 2nd ed. Lilongwe: Ministry of from Mussa et al. (2017) that a larger number Agriculture and Food Security. (65.7%) of women from Malawi are involved in the Haambiya L, Matiya G, Msukwa A, Kapute F, Sikawa D. informal cross border fish trade. The issue of women 2013. Factors that influence effectiveness of local empowerment is therefore, crucial as 70% of women organisations in promoting community participation in the Southern African Development Community in artisanal fisheries management in Malawi: A case (SADC) are involved in the informal cross border of Mbenji island fishery. Malawi Aqua Fish. 2(1):1-7. trade (Mussa et al. 2017). Jayalal L, Ramachandran A. 2012. Export trend of Indian The study recommends that the Government of ornamental fish industry. Agric Bio J North America. 3(11):439–451. Malawi should make a balanced decision on policy doi:10.5251/abjna.2012.3.11.439.451. issues regarding regulation of fish exports. This Kapute F. 2009. Fish quality and processing in Malawi: comes at a time when demand for fish on the local Responding to challenges through institutional market in the country is not satisfied. Further, the capacity building. Reykjavik: United Nations government should consider exploring fish University Fisheries Training Programme (UNU- processing methods that ensure highest quality for FTP) Project Report. exports to highly regulated markets. The more Kapute F. 2017. Chasing Malawi fish: the irony of local women (86.2%) than men (13.8%) who exported fish fish scarcity versus exports. The 2017 Malawi nd th from Blantyre District provides insights on the Fisheries and Aquaculture Symposium 22 to 24 unsustainability of fish exports. Therefore, a policy November 2017. Sun n Sand Holiday Resort, Mangochi. Malawi. that considers mainstreaming of gender into the Manyungwa-Pasani CL, Hara M, Chimatiro SK. 2017. policy may ensure the sustainability of livelihoods of Women’s participation in fish value chains and value women in fish exports. chain governance in Malawi: a case of Msaka (Lake Malawi) and Kachulu (Lake Chilwa). Cape Town: Acknowledgements Institute for Poverty, Land and Agrarian Studies. We thank the Malawi Government’s Department Working Paper No.: 45. of Fisheries and Blantyre District Fisheries Office for Mussa H, Kaunda E, Chimatiro S, Banda L, Nankwenya providing the data used in this study. B, Nyengere J. 2017. Assessment of informal cross border fish trade in the southern African region: a case of Malawi and Zambia. Paper presented at: Malawi References Fisheries and Aquaculture Forum; Mangochi, Malawi. Bhattacherjee A. 2012. Social science research: principles, Singini W. 2013. Bioeconomic analysis of Chambo methods and practices. 2nd ed. Florida: Anol (Oreochromis spp.) and Kambuzi (small Bhattacherjee 159 p. Haplochromine spp.) fish stocks of Lake Malombe. Commonwealth/GTZ. 2007. ESA meeting on trade and [PhD Thesis], University of Malawi. 199 p. sustainable approaches to fisheries negotiations under Ukaonu SU, Mbawuike BC, Uluwajoba EO, Williams AB, WTO/EPA. Port Louis: ESA. Report No.: (07) 05. Ajuonu N, Omogoriota HO, Olakolu FC, Adegbile Donda S, Njaya F. 2007. Food security and poverty OM, Myade EF. 2011. Volume and value of alleviation through improved valuation and ornamental fishes in the Nigerian export trade. Agr governance of river fisheries in Africa: Fisheries co- Bio J North America. 2(4):661-664. management in Malawi, analysis of the underlying doi:10.5251/abjna.2011.2.4.661.664.